Title: Affymetrix CEL analysis. Description:
nucleic acid hybridization to array protocol
P-AFFY-3 GeneChip hybridization
normalization data transformation protocol
chips were normalized together using robust multichip average. Median polish was used for the probe level summary.
nucleic acid labeling protocol
Affymstrix GeneChip IVT Labeling Kit
nucleic acid extraction protocol
Total RNA was extracted from leaf using Qiagen Plant Mini Kit.
Heat stress treatment protocol
Thirty days after sowing, switchgrass seedlings were subjected to heat stress treatment of 38 degrees /30 degrees C (day/night) for 50 days.
Switchgrass (Panicum virgatum L.) Alamo cultivar seeds were sown in pots (0.2 meter diameter ×0.45m tall) filled with pure, fine sand soil and grown in growth chambers (Conviron Ltd., Winnipeg, Canada) at 28 degrees /20 degrees C (day/night) with a photoperiod of 14 h/10h (day/night). After germination, switchgrass seedlings were thinned to four plants per pot. Leaf samples from plants growing under control condition (28 degrees /20 degrees C) and heat stress (38 degrees /30 degrees C) were harvested and snap frozen in liquid nitrogen for RNA isolation at the end of treatment.