Title: Affymetrix CEL analysis. Description:
Affymetrix Generic Hybridization
The plates were kept in a growth chamber with a conditions; 23Â°C, 16 hr light and 8 hr dark cycles
First-strand of cDNA was synthesized using 16 ¼g of total RNA and SuperScript Double-stranded cDNA Synthesis kit (Invitrogen). The labeled cRNA was synthesized using BioArray RNA Transcripts Labeling Kit (T7) (ENZO Life Sciences inc.). After labeling, the cRNA was digested partially by incubating in 40 mM Tris-acetate, pH8.0, 100 mM KOAc, 30 mM MgOAc, at 94Â°C for 35 min. Then 30 ¼g cRNA was used for microarrary hybridization.
After stratification at 4Â°C for 4 days, seeds were allowed to germinate on the MS-2% sucrose plate containing 0.5 uM ABA for two days
After stratification at 4Â°C for 4 days, seeds were allowed to germinate on the MS-2% sucrose plate for two days
Total RNA was extracted from frozen tissue using TRIzol reagent (Invitrogen) and purified using a RNease MinElute Cleanup Kit (Qiagen).