Transcriptional profiles of drought-responsive genes in modulating transcription, signal transduction and biochemical pathways in tomato
Hybridization: IL2-5 drought stress/control |
Hybridization protocol |
Hybridization solution | N/A |
Solution concentration | N/A |
Blocking agent | N/A |
Wash procedure | the arrays were washed with 0.2% SDS, 2Ă—SSC at 42oC for 5 min, followed by washing with 0.2% SSC for 5 min at room temperature. |
Labeled Target Quantity | N/A |
Hybridization Time | overnight |
Hybridization Volume | N/A |
Hybridization Temperature | 42oC |
Hybridization Instrument | N/A |
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Scan information |
Scan hardware | LuxScan 10KA confocal laser scanner |
scan software | N/A |
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Image analysis information |
Software | ImaGene ver 5.5.4 |
Analysis algorithm | 1) Images Cy5 = Red Cy3 = Green
2) Grid Flexibility Enforce grid constraints = yes Local flexibility = 6.0 pixels Flexibility = 75%
3) Quality Control Empty spots = no Poor spots = yes Find negative spots = no
4) Measurements Background buffer = 2.25 Background width = 5.0 Auto seqmentation = no Signal low cutoff = 20% Signal high cutoff = 100% Background low cutoff = 0% Background high cutoff = 88%
5) Poor Spot Parameters Contamination of Background/Signal = no Ignored pixels % = yes (setting = 69) Open perimeter = no Shape regularity = no Area to perimeter = yes (setting = 0.6) Off-set value = yes (setting = 17.5) |
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Normalization information |
Normalization software | marray package under R |
Normalization algorithm | Print-tip specific Lowess Normalization |
Normalization strategy | Total Array |
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