RNA-Seq of ovis aries: adult female cinical mastitic mammary tissue
mRNA isolation and fragmentation were conducted using NEBNextR Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, Inc.) and fragmentation buffer, respectively. We used reverse transcriptase and random hexamer-primer to synthesize first-strand cDNA with cleaved short RNA fragments as template. We subsequently synthesized second-strand cDNA by adding reaction buffer, dNTPs, RNase H, and DNA polymerase I using first-strand cDNA as template. The double-stranded cDNA was ultimately purified by end repair, adaptor ligationgel, selection, and library enrichment successively. The quality assessment and quantification of the sample libraries were performed using QubitR Fluorometers (Thermo Fisher Scientific) and KAPA Library Quantification Kit (KAPA Biosystems), respectively. The resulting high-quality libraries were sequenced in parallel on Illumina HiSeq TM 2000 system