Title: Affymetrix CEL analysis. Description:
Affymetrix Generic Hybridization
nucleic acid labeling protocol
Labeling of samples was carried out according to the manufacturer’s standard protocol
Background correction,quantile normalization, and gene expression analysis were performed using RMA (Bolstad, B.M., Irizarry R. A., Astrand, M., and Speed, T.P. (2003)A Comparison of Normalization Methods for High Density Oligonucleotide Array Data Based on Bias and Variance. Bioinformatics 19(2):185-193).
Primary dermal fibroblast cultures from individuals 915-07 and 909-6 were established from skin biopsies by routine procedures. Control skin biopsy samples were obtained from normal tissue from patients of similar age and sex undergoing dermatologic procedures. All fibroblasts were used at the same passage number. Fibroblasts were routinely maintained in DMEM (PAA, Somerset) in the presence of 10% foetal calf serum (PAA, Somerset) at 37°C, in 5% CO2. Cells were harvested by trypsin treatment. Fibroblasts were expanded until full confluency was achieved.
(Parameters: time unit = seconds, temperature unit = C, media = DMEM)
Fibroblasts were expanded until full confluency was achieved. Cells were harvested by trypsin treatment. Total RNA was extracted from cell lines using RNeasy Mini Kit and QIA Shedder (Qiagen, USA).
(Parameters: Extracted product = total_RNA, Amplification = none)