Title: Affymetrix CEL analysis. Description:
Affymetrix Generic Hybridization
RNA was isolated using the RNeasy Plant Mini Protocol (QIAGEN, Clifton Hill, Australia) and the quality of the RNA was assessed using the Agilent Bioanalyzer (Agilent Technologies, Palo Alto, CA) where spectrophotometric analysis was carried out to determine the A260 to A280 ratio and the concentration. For qRT-PCR, cDNA synthesis was carried out using the Biorad (Perth) iScript cDNA synthesis kit according to manufacturers instructions. For arrays, the cRNA was prepared from 3 ug of total RNA almost exactly as described in the Affymetrix GeneChip Expression Analysis Technical Manual.
nucleic acid extraction protocol
Standard procedures were used to obtain total RNA (Qiagen) from the epididymal white adipose tissue of 15 day old wt and Ercc1-/- animals (n=4).
(Parameters: Extracted product = total_RNA, Amplification = none)
Epididymal white adipose tissue was dissected from 15 day old wt and Ercc1-/- animals (n=4).
(Parameters: time unit = seconds, temperature unit = C)