Title: Affymetrix CEL analysis. Description:
Expression Analysis Technical Manual, with Specific Protocols for Use with the Hybridization, Wash, and Stain Kit. P/N 702232 Rev. 3 http://media.affymetrix.com/support/downloads/manuals/expression_analysis_technical_manual.pdf (Parameters: Chamber type = Affymetrix- GeneChip Hyb Oven 640, Quantity of label target used = 10, Mass unit = Micro gram, time = 16, Tiny time unit = hours, Volume = 200, Volume unit = Micro litre, temperature = 45)
Affymetrix GeneChip Whole Transcript (WT)Sense Target Labeling Assay P/N 701880 Rev. 5 (Parameters: Amount of nucleic acid labeled = 250, Amplification = none, Mass unit = Nano gram)
Infection by retroviral-mediated gene transfer with either pBABE-puro-HA-PIASy or empty vector were performed as previously described (Bischof et al., 2006).
Total RNA was extracted using Rneasy Kit according to the manufacturer’s recommendations (Qiagen). (Parameters: Extracted product = total_RNA, Amplification = none)
Culturing of human diploid WI38 fibroblasts (ATCC) was performed at physiological oxygen concentration of 3% as previously described (Bischof et al., 2006). Parameters: time unit = seconds, temperature unit = C)