RNA-Seq of Bos taurus: adult ovarian follicular cells
RNA sequencing was performed using the TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA) following manufacturer's instructions. Briefly, 2 g of total RNA from each GC sample was used for library preparation. The RNA concentration and purity were measured using NanoDropTM (Thermos Scientific, Waltham, MA) and integrity was assessed by Bioanalyzer (Agilent, Santa Clara, CA). The mRNA was first enriched from total RNA using oligo dT magnetic beads and then poly(A) RNA was fragmented and cDNA synthesized. Final cDNA repair was then performed. The 3' ends were adenylated, and universal barcode adapters were ligated to the cDNA fragments to perform solid phase PCR and produce the sequencing library; the latter was evaluated and quantified using an Agilent 2100 Bioanalyzer and quantitative PCR with the KAPA Library Quantification kit (KAPA Biosystems, Foster City, CA, USA). Finally, libraries were pooled to perform multiplexing sequencing using HiSeq SBS v4 High Output Kit. Cluster generation and sequencing were performed on the Illumina HiSeq 2500, with 2  125 bp pair-end reads produced.