A tomato fruit RNA extraction method that isolates mRNAs encoding secreted and endomembrane associated proteins

Hybridization: Ailsa Craig

Hybridization protocol
Hybridization solution1X hybridization buffer
Solution concentration100 mM MES, 1 M NaCl, 20 mM EDTA, 0.01% Tween 20
Blocking agentHerring sperm DNA (final concentration 100ng)
Wash procedureAutomated fluidics station consisting of wash buffers A and B. Wash buffer A is a non-stringent buffer containing 6X SSPE, 0.01% Tween 20. Wash buffer B is a stringent buffer containing 100 mM MES, 0.1 M NaCl, 0.01% Tween 20
Labeled Target Quantity5 ug
Hybridization Time16 hours
Hybridization Volume80 ul
Hybridization Temperature45C
Hybridization InstrumentHybridization oven 640 (Affymetrix)

Scan information
Scan hardwareHewlett-Packard GeneArray Scanner G2500A
scan softwareN/A

Image analysis information
SoftwareAffymetrix GeneChip Operating Software (GCOS)
Analysis algorithmMAS 5 software

Normalization information
Normalization softwareRMA
Normalization algorithmN/A
Normalization strategyTotal array