RNA was isolated from the milk fat globule using Trizol following the manufacturer's protocol
Label
Biotin
Label protocol
440 ng total RNA was used for in vitro transcription, overnight for 14h using biotin-11-dUTP for labeling of the cRNA product using the Illumina®TotalPrep RNA amplification kit (Ambion, Inc., Austin, TX)
Hybridization protocol
750 ng of labeled cRNAs were hybridized to the Human Ref 8 V2 Beadchip arrays (Illumina Inc., San Diego, CA) at 55oC overnight (16h) following the Illumina Whole-Genome Gene Expression Protocol for BeadStation (Doc. # 111226048 Rev. B, Illumina Inc) and stained with 1 ug/ml streptavidin-Cy3 (Amersham Biosciences, Piscataway, NJ) for visualization
Scan protocol
Probe arrays were scanned using the BeadArray reader (Factor 1)
Description
Day4_GH3_2PM
Data processing
Gene expression analysis was performed using the Sentrix BeadChip and BeadStation system from Illumina, Inc
