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Experiment: Systemic responses to Pseudomonas syringae infection

Experiment Description

NASCArrays Experiment Reference Number: NASCARRAYS-437

We used three different strains of Pseudomonas syringae pv tomato DC3000 to investigate systemic responses to infection in Arabidopsis and the development of SAR. Wildtype DC3000, the hrpA mutant and DC3000 carrying the avirulence gene avrRpm1 were syringe infiltrated into 4 week old plants at a concentration of 10e8 cfu/ml. At least 5 leaves per plant were infiltrated and at least 10 plants were pooled for each sample. Systemic, uninfected tissue was then harvested at 8, 12 and 21h after inoculation. Three independent experiments were carried out to give three biological replicates for each timepoint.

About the Experimenter

Name:Dr William Truman
Head of Lab Name:Prof Murray Grant
Lab:
Address:School of Biosciences
University of Exeter
Geoffrey Pope Building
Stocker Road
Exeter
Postcode: EX44QD
Country: UK
 
Telephone Number: 01392269162

All of the data available in this website/database is free, and you are free to do whatever you please with it. If you intend to publish work based on any of this data, please acknowledge us, contact the experimenter above, and either acknowledge them or use them as co-authors in the work.

About this Experiment

Experiment Type: time_series_design; pathogenicity_design
Number of Slides:27
 
Experimental Parameters:
parameterinfect
parametertimepoint
Quality Control Measures Taken:
no-plants-pooled5 plants
References:
 
Other Information:

Slides in this Experiment

Hybridisation Set: Truman: Systemic responses to Pseudomonas syringae infection_genome

Slide: Truman_2-1_8h-hrpA_Rep1_ATH1

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-1_8h-hrpA_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-1_8h-hrpA_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
Tissue: leave
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.82011526823
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.39
NoiseAvg:2.50,Stdev:0.08,Max:2.9,Min:2.2
Central-Avg:5619,Count:9
Corner+Avg:79,Count:32
Corner-Avg:7529,Count:32
BackgroundAvg:54.88,Stdev:0.30,Max:55.5,Min:54.0
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.820115268230
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-2_8h-DC3000_Rep1_ATH1

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-2_8h-DC3000_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-2_8h-DC3000_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.858121871948
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.22
NoiseAvg:2.32,Stdev:0.07,Max:2.6,Min:2.2
Central-Avg:5392,Count:9
Corner+Avg:76,Count:32
Corner-Avg:7092,Count:32
BackgroundAvg:52.33,Stdev:0.88,Max:54.7,Min:50.4
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.858121871948
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-3_8h-avrRpm1_Rep1_ATH1

Add to Slide Selection

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-3_8h-avrRpm1_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-3_8h-avrRpm1_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.953052282333
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.49
NoiseAvg:2.48,Stdev:0.11,Max:2.8,Min:2.2
Central-Avg:5099,Count:9
Corner+Avg:69,Count:32
Corner-Avg:6735,Count:32
BackgroundAvg:59.79,Stdev:1.07,Max:62.8,Min:57.7
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.953052282333
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-4_12h-hrpA_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-4_12h-hrpA_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-4_12h-hrpA_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.960087716579
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.55
NoiseAvg:2.52,Stdev:0.05,Max:2.7,Min:2.3
Central-Avg:4974,Count:9
Corner+Avg:72,Count:32
Corner-Avg:7361,Count:32
BackgroundAvg:58.59,Stdev:0.43,Max:59.9,Min:57.3
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.960087716579
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-5_12h-DC3000_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-5_12h-DC3000_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-5_12h-DC3000_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.012127995491
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.23
NoiseAvg:2.22,Stdev:0.06,Max:2.6,Min:2.1
Central-Avg:4214,Count:9
Corner+Avg:68,Count:32
Corner-Avg:6308,Count:32
BackgroundAvg:52.64,Stdev:0.32,Max:53.6,Min:51.7
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.012127995491
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-6_12h-avrRpm1_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-6_12h-avrRpm1_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-6_12h-avrRpm1_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.048158884048
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.30
NoiseAvg:2.19,Stdev:0.05,Max:2.3,Min:2.0
Central-Avg:3851,Count:9
Corner+Avg:64,Count:32
Corner-Avg:5914,Count:32
BackgroundAvg:54.42,Stdev:0.20,Max:54.9,Min:54.0
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.048158884048
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-7_21h-hrpA_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-7_21h-hrpA_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-7_21h-hrpA_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.829517066479
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.49
NoiseAvg:2.49,Stdev:0.07,Max:2.7,Min:2.2
Central-Avg:4963,Count:9
Corner+Avg:70,Count:32
Corner-Avg:6844,Count:32
BackgroundAvg:58.71,Stdev:0.64,Max:60.9,Min:56.9
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.829517066479
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-8_21h-DC3000_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-8_21h-DC3000_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-8_21h-DC3000_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.037037372589
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.41
NoiseAvg:2.34,Stdev:0.12,Max:2.9,Min:2.0
Central-Avg:4950,Count:9
Corner+Avg:67,Count:32
Corner-Avg:6497,Count:32
BackgroundAvg:57.46,Stdev:0.68,Max:59.2,Min:56.1
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.037037372589
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-9_21h-avrRpm1_Rep1_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-9_21h-avrRpm1_Rep1_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-9_21h-avrRpm1_Rep1_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.983953237534
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.37
NoiseAvg:2.46,Stdev:0.14,Max:2.9,Min:2.2
Central-Avg:4805,Count:9
Corner+Avg:64,Count:32
Corner-Avg:6316,Count:32
BackgroundAvg:57.58,Stdev:1.70,Max:63.4,Min:54.6
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.983953237534
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-10_8h-hrpA_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-10_8h-hrpA_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-10_8h-hrpA_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.128173828125
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.18
NoiseAvg:2.07,Stdev:0.08,Max:2.5,Min:1.9
Central-Avg:4166,Count:9
Corner+Avg:60,Count:32
Corner-Avg:5543,Count:32
BackgroundAvg:51.49,Stdev:0.69,Max:55.1,Min:50.8
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.128173828125
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-11_8h-DC3000_Rep2_ATH1

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-11_8h-DC3000_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-11_8h-DC3000_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.930367767811
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.36
NoiseAvg:2.31,Stdev:0.06,Max:2.5,Min:2.2
Central-Avg:4851,Count:9
Corner+Avg:68,Count:32
Corner-Avg:6602,Count:32
BackgroundAvg:58.44,Stdev:0.34,Max:59.1,Min:57.1
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.930367767811
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-12_8h-avrRpm1_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-12_8h-avrRpm1_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-12_8h-avrRpm1_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.74541670084
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.41
NoiseAvg:2.58,Stdev:0.08,Max:2.8,Min:2.3
Central-Avg:5142,Count:9
Corner+Avg:72,Count:32
Corner-Avg:7069,Count:32
BackgroundAvg:61.48,Stdev:0.95,Max:63.4,Min:59.0
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.745416700840
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-13_12h-hrpA_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-13_12h-hrpA_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-13_12h-hrpA_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.743233561516
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.54
NoiseAvg:2.55,Stdev:0.07,Max:2.8,Min:2.4
Central-Avg:5809,Count:9
Corner+Avg:81,Count:32
Corner-Avg:7966,Count:32
BackgroundAvg:59.97,Stdev:0.52,Max:61.1,Min:58.5
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.743233561516
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-14_12h-DC3000_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-14_12h-DC3000_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-14_12h-DC3000_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.031928300858
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.08
NoiseAvg:1.90,Stdev:0.04,Max:2.0,Min:1.8
Central-Avg:4898,Count:9
Corner+Avg:75,Count:32
Corner-Avg:7783,Count:32
BackgroundAvg:47.25,Stdev:0.28,Max:48.0,Min:46.4
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.031928300858
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-15_12h-avrRpm1_Rep2_ATH1

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-15_12h-avrRpm1_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-15_12h-avrRpm1_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.682912826538
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.48
NoiseAvg:2.63,Stdev:0.06,Max:2.8,Min:2.5
Central-Avg:5408,Count:9
Corner+Avg:78,Count:32
Corner-Avg:7330,Count:32
BackgroundAvg:60.69,Stdev:0.36,Max:61.5,Min:59.5
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.682912826538
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-16_21h-hrpA_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-16_21h-hrpA_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-16_21h-hrpA_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.718620657921
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.62
NoiseAvg:2.69,Stdev:0.06,Max:2.9,Min:2.4
Central-Avg:6555,Count:9
Corner+Avg:82,Count:32
Corner-Avg:8085,Count:32
BackgroundAvg:64.22,Stdev:0.40,Max:65.1,Min:62.9
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.718620657921
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-17_21h-DC3000_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-17_21h-DC3000_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-17_21h-DC3000_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.465970516205
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.23
NoiseAvg:2.07,Stdev:0.08,Max:2.4,Min:1.8
Central-Avg:4569,Count:9
Corner+Avg:64,Count:32
Corner-Avg:6693,Count:32
BackgroundAvg:53.11,Stdev:0.64,Max:54.8,Min:51.4
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.465970516205
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-18_21h-avrRpm1_Rep2_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-18_21h-avrRpm1_Rep2_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-18_21h-avrRpm1_Rep2_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.42604637146
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.23
NoiseAvg:2.18,Stdev:0.06,Max:2.4,Min:2.0
Central-Avg:4601,Count:9
Corner+Avg:61,Count:32
Corner-Avg:6104,Count:32
BackgroundAvg:56.53,Stdev:0.64,Max:57.8,Min:54.8
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.426046371460
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-19_8h-hrpA_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-19_8h-hrpA_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-19_8h-hrpA_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.238400816917
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.37
NoiseAvg:2.29,Stdev:0.06,Max:2.6,Min:2.2
Central-Avg:5161,Count:9
Corner+Avg:65,Count:32
Corner-Avg:6880,Count:32
BackgroundAvg:57.04,Stdev:0.37,Max:57.7,Min:55.7
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.238400816917
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-20_8h-DC3000_Rep3_ATH1

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-20_8h-DC3000_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-20_8h-DC3000_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.934285521507
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.44
NoiseAvg:2.38,Stdev:0.08,Max:2.6,Min:2.2
Central-Avg:6285,Count:9
Corner+Avg:74,Count:32
Corner-Avg:7927,Count:32
BackgroundAvg:59.53,Stdev:0.88,Max:62.4,Min:57.5
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.934285521507
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-21_8h-avrRpm1_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-21_8h-avrRpm1_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-21_8h-avrRpm1_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 8h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.603649735451
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.39
NoiseAvg:2.22,Stdev:0.07,Max:2.4,Min:2.1
Central-Avg:4176,Count:9
Corner+Avg:54,Count:32
Corner-Avg:5536,Count:32
BackgroundAvg:57.92,Stdev:1.06,Max:62.1,Min:56.2
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.603649735451
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-22_12h-hrpA_Rep3_ATH1

Add to Slide Selection

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-22_12h-hrpA_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-22_12h-hrpA_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.09977543354
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.36
NoiseAvg:2.30,Stdev:0.06,Max:2.6,Min:2.1
Central-Avg:5166,Count:9
Corner+Avg:65,Count:32
Corner-Avg:6746,Count:32
BackgroundAvg:56.01,Stdev:0.66,Max:57.9,Min:54.9
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.099775433540
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-23_12h-DC3000_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-23_12h-DC3000_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-23_12h-DC3000_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.169638395309
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.17
NoiseAvg:2.09,Stdev:0.05,Max:2.2,Min:1.9
Central-Avg:4521,Count:9
Corner+Avg:62,Count:32
Corner-Avg:6385,Count:32
BackgroundAvg:52.55,Stdev:0.32,Max:53.2,Min:51.6
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.169638395309
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-24_12h-avrRpm1_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-24_12h-avrRpm1_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-24_12h-avrRpm1_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 12h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:1.140247344971
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.29
NoiseAvg:2.25,Stdev:0.10,Max:2.7,Min:2.0
Central-Avg:6084,Count:9
Corner+Avg:75,Count:32
Corner-Avg:7409,Count:32
BackgroundAvg:57.18,Stdev:0.96,Max:60.8,Min:55.7
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF1.140247344971
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-25_21h-hrpA_Rep3_ATH1

Add to Slide Selection

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Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-25_21h-hrpA_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-25_21h-hrpA_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.518167436123
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.43
NoiseAvg:2.71,Stdev:0.07,Max:2.9,Min:2.5
Central-Avg:4683,Count:9
Corner+Avg:75,Count:32
Corner-Avg:6123,Count:32
BackgroundAvg:58.87,Stdev:0.53,Max:60.6,Min:57.8
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.518167436123
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-26_21h-DC3000_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-26_21h-DC3000_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-26_21h-DC3000_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.797910928726
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ2.09
NoiseAvg:2.03,Stdev:0.06,Max:2.2,Min:1.9
Central-Avg:4881,Count:9
Corner+Avg:61,Count:32
Corner-Avg:5963,Count:32
BackgroundAvg:49.44,Stdev:0.20,Max:50.2,Min:48.9
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.797910928726
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015

Slide: Truman_2-27_21h-avrRpm1_Rep3_ATH1

Add to Slide Selection

Get the data!

Summary BioSource 1 Information Extraction / Labelling Protocol for Source 1 Information Slide Information Hybridisation Analysis Information for "Affymetrix MAS 5.0 Scaling Protocol" normalised data.

Summary Information

This slide has had 1 extract(s) hybridised to it (1-colour).

  • Extract 1 ("Truman_2-27_21h-avrRpm1_Rep3_ATH1") was made from "Arabidopsis thaliana" and labelled using "Biotin Labelled cRNA using Affymetrix Enzo Kit".

The slide used was "Affymetrix ATH1 Arabidopsis Genome Array".

Click on the options above for more information about this slide and its preparation.

Hybridisation Protocol Information

Name of Protocol: Affymetrix Antibody Amplification Protocol
Solution:
BufferEDTA, NaCl, Tween20 according to Affymetrix protocol
Water
Blocking Agent:
Herring Sperm DNA(Source: Promega)100pg/ml
Acetylated BSA(Source: Sigma-Aldrich)0.1mg/ml
Wash Procedure:
Step 1(Source: Affymetrix)10 cycles of 2 mixes/cycle with Non-Stringent buffer
Step 2(Source: Affymetrix)4 cycles of 15 mixes/cycle with Stringent buffer
Step 3(Source: Affymetrix)Stain the probe array for 10 minutes in Streptavaidin Phycoerythrin (SAPE) solution at 25 C
Step 4(Source: Affymetrix)10 cycles of 4 mixes/cycle with Non-Stringent buffer
Step 5(Source: Affymetrix)Stain the probe array for 10 minutes in antibody solution
Step 6(Source: Affymetrix)Stain the probe array for 10 minutes in SAPE solution at 25 C
Step 7(Source: Affymetrix)15 cycles of 4mixes/cycle with Non-Stringent buffer at 30 C
Step 8(Source: Affymetrix)Hold at 25 C
Quantity Used:
Amount15 microgram Fragmented cRNA
Time: 16 hours
Concentration: 50 pg/ml
Volume: 200 microlitres
Temperature: 45 C
Other Information:
Spike(Source: Affymetrix)Control Oligo B2 to 50pM for landing lights
Spike(Source: Affymetrix)20X Eukaryotic Hybridisation Controls (bioB (1.5pM, bioC 5pM, bioD 25pM, cre 100pM)
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

BioSource 1 Information

BioSource Name: Truman_2-27_21h-avrRpm1_Rep3_ATH1
Organism: Arabidopsis thaliana
Alias: Col-5
Stock Code: N1688
Age: 21h
Growth Conditions:
LocationGrowth Room
Growth substrate(Source: Commercial soil)Levingtons F2 compost (Fisons, Ipswich, UK)
stratification2 days in fridge at 4oC
Growth protocolA. thaliana was sown in Levingtons F2 compost and vernalised for 2 days at 4°C. Plants were grown under short day conditions in a controlled environment chamber (10 h light, 100?150 µEinsteins at 22°C day, 20°C night) for 4-5 weeks before use.
Developmental Stage:
developmental-stage(Source: Boyes et al., Plant Cell 2001 13, 1499)3.90
in vivo Treatment:
treatmentLeaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample.
Additional Organism Information:
sample descriptionSystemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested
Other Information:

Protocols for BioSource 1

Extraction Protocol Information

Name of Protocol: Qiagen / Trizol
Method:
ProtocolQiagen RNAeasy kit protocol
Type: total RNA
Amplification: Amplified during labelling process - see labelling protocol
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Labelling Protocol Information

Name of Protocol: Biotin Labelled cRNA using Affymetrix Enzo Kit
Amount Labelled: Unknown
Label Name: Biotin
Labelling Method: In-vitro transcription method
Other Information:
ProtocolSourceAffymetrix GeneChip Technical Analysis Manual

Slide Information

Date Hybridised:

Array Design Information

Slide Design Name: Affymetrix ATH1 Arabidopsis Genome Array
Array Type: Affymetrix Gene Chip
Number of Spots on the Array: 22810

Scanning Protocol Information

Name: Affymetrix MAS 5.0 Standard Scanning
Width of Image Produced (Pixels): 4733
Height of Image Produced (Pixels): 4733
X Resolution (microns): 3
Y Resolution (microns): 3
Scanner Name: Agilent 2500A GeneArray Scanner
Software Name: Affymetrix Micro Array Suite 5.0

Image Analysis Information

Scaling Factor:0.92348241806
Name: Affymetrix MAS 5.0 Standard Image Analysis
Software Name: Affymetrix Microarray Analysis Suite 5.0

Normalisation Information

Strategy Name: Affymetrix MAS 5.0 Scaling Protocol
Algorithm: The top 2% and bottom 2% of signal intensities are excluded, then the mean is calculated. The original signal values are scaled such that the mean is made equal to 100.
Control Elements:
none
Normalisation Protocol Parameters:
RawQ1.97
NoiseAvg:1.99,Stdev:0.05,Max:2.2,Min:1.9
Central-Avg:4417,Count:9
Corner+Avg:57,Count:32
Corner-Avg:5572,Count:32
BackgroundAvg:49.04,Stdev:0.25,Max:49.8,Min:48.5
Gamma1H0.0025
VZ4
Gamma2L0.003
Epsilon0.5
TGT100
Perturbation1.1
Alpha20.06
BF
BG2
Gamma2H0.003
SF0.923482418060
Alpha10.04
SFGeneAll
HZ4
NF1.000000000000
SmoothFactorBG100
Gamma1L0.0025
Tau0.015


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