7 protocols
AccessionType
feature_extraction
Title: Affymetrix CEL analysis. Description:
hybridization protocol
Affymetrix Generic Hybridization
grow
The plates were kept in a growth chamber with a conditions; 23°C, 16 hr light and 8 hr dark cycles
labeling
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 94°C for 35 min. Then 30 ¼g cRNA was used for microarrary hybridization.
specified_biomaterial_action
After stratification at 4°C for 4 days, seeds were allowed to germinate on the MS-2% sucrose plate containing 0.5 uM ABA for two days
specified_biomaterial_action
After stratification at 4°C for 4 days, seeds were allowed to germinate on the MS-2% sucrose plate for two days
nucleic_acid_extraction
Total RNA was extracted from frozen tissue using TRIzol reagent (Invitrogen) and purified using a RNease MinElute Cleanup Kit (Qiagen).