Total RNA was isolated from intestinal tissue using the RNeasy lipid tissue extraction kit and on-column DNase treatment following the manufacturer’s recommendations (Qiagen, Valencia, USA). RNA concentration and quality were respectively assessed using the NanoDrop spectrophotometer (Thermo Scientific, Wilmington, USA) and the Agilent Technologies 2100 bioanalyzer (Agilent, Santa Clara, USA). Only total RNA samples with a RIN higher than seven were included in the following experiments.
Label
biotin
Label protocol
biotin labeling were performed using the TargetAmp Labeling Kit (Illumina, San Diego, USA)
Hybridization protocol
Hybridization of 700 ng of each biotinylated cRNA sample on the BeadChip was performed at 58°C for 16–18 hours
Scan protocol
BeadChips were scanned using the Illumina Bead Array Reader (Illumina, San Diego, USA).
Description
2102
Data processing
raw data were log2 transformed and quantile normalized using the lumi package from bioconductor in the R statistical software
