title: RNA-seq buffy coat (white blood cells) Bovine (Holstein cows)
design: Total RNA from WBC was extracted using the TRIzol Reagent (Invitrogen, CA, USA) following the manufacturer's protocol. RNA concentrations and quality were assessed using a Fragment Analyzer instrument (DNF-472, Advanced Analytical Technologies, Ames, IA). Total RNA samples were sent to BGI Genomics (Beijing, China) for library preparation and sequencing. PolyA-enriched RNA libraries were constructed using an MGIEasy RNA Library Prep Set (BGI, Beijing, China) according to the manufacturers protocol. Briefly, mRNA was purified using oligo (dT) attached magnetic beads, and then fragmented into small pieces. Following first- and second- strands cDNA synthesis, double-stranded cDNA fragments were subjected to end-repair, adenylation, adaptor ligation, and PCR amplification. Each purified library was assessed using the Fragment Analyzer, quantified with a fluorometer (Qubit), and sequenced on a MGI DNBSEQ G400 instrument resulting in paired-end reads of 150 bp and an average read count of 67 million paired reads per sample.