Total RNA from buccal scrapings was isolated using miRNeasy Mini Kit (Qiagen, Valencia, CA)
Label
biotin
Label protocol
Biotin labeling was performed using the Ambion WT Expression Kit (Life Technologies, Grand Island, NY) according to the manufacturer's protocol, followed by the GeneChip WT Terminal Labeling and Controls Kit (Affymetrix, Santa Clara, CA).
Hybridization protocol
The labeled, fragmented DNA was hybridized to the GeneChip Human Gene 1.0 ST Array (Affymetrix, Santa Clara, CA) for 18 hours in a GeneChip Hybridization oven at 640 at 45 deg. C with rotation (60 rpm). The hybridized samples were washed and stained using an Affymetrix fluidics station 450 with streptavidin-R-phycoerythrin (SAPE) and the signal was amplified using a biotinylated goat anti-streptavidin antibody followed by another SAPE staining (Hybridization, Washing and Staining Kit, Affymetrix, Santa Clara, CA).
Scan protocol
After staining, microarrays were immediately scanned using an Affymetrix GeneArray Scanner 3000 7G Plus (Affymetrix, Santa Clara, CA).
Description
CQ 1361 0252 buccal epithelial RNA from nonsmoking, healthy female exposed to smokeless-coal
Data processing
RMA normalization was conducted using custom CDF: HuGene10stv1_Hs_ENTREZG.cdf [Brainarray Version 14]
