Instrument: Illumina Genome Analyzer IIx
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Tail vein blood was collected from 40 Holstein cows at the University of Wisconsin Dairy Cattle Center, 7 Jersey cows at the University of Wisconsin Arlington Agricultural Research Station, and 45 Cholistani cows at GujaitPeer Farm, Bahawalpur, Punjab, Pakistan. Immediately following blood sampling, equal volumes of whole blood were pooled before white blood cells were isolated by centrifugation at 3000 g for 15 min. The white blood cells were washed twice with red blood cell lysis buffer, followed by RNA extraction using the Qiagen RNAEasy Mini Kit (Qiagen, Valencia, CA). For the Cholistani breed, RNA was transported in RNAStable (Biomatrica, San Diego, CA), a storage matrix mimicking an anhydrobiotic environment, to preserve its integrity. We quality checked all RNA samples using Agilent Bioanalyzer 2100 (Agilent, Santa Clara CA) before generation of sequencing libraries. Polyadenylated RNA was selected using magnetic oligo dT beads and subjected to Illumina's mRNA-Seq library generation.