array scanning and feature extraction protocol
Title: Affymetrix CEL analysis. Description:
Title: Affymetrix CHP Analysis (ExpressionStat). Description:
Approximately 100 ng of RNAwas used as template for the synthesis of labeled cRNA probe using the Two Cycle Amplification Kit (Affymetrix). Briefly, the first cycle of cDNA synthesis was carried out using an oligo (dT) primer fused to a T7 RNA polymerase promoter and was followed by the first cycle of in vitro transcription using T7 RNA polymerase to generate cRNA. The cRNA was used as a template for the second cycle of cDNA synthesis followed by the second cycle of in vitro transcription, to generate the biotin-labeled antisense cRNA.
The seeds were dissected out from siliques in RNAlater solution (Ambion) and total RNAs were extracted using the RNeasy Plant Mini Kit (QIAGEN). RNA quality and integrity were determined using an Agilent 2100 Bioanalyzer (Agilent Technologies).
Plants were grown on soil under constant illumination.