title: GSM6745531: granulosa cells with INHBA siRNA transfected  in vitro,rep 1; Ovis aries; RNA-Seq
library_construction_protocol: TRIZOL was used to extract total RNA and the concentration and purity of RNA were detected by the Nanodrop 2000 spectrophotometer. The eukaryotic mRNA was enriched with magnetic beads with Oligo (dT), and the mRNA was randomly interrupted by adding Fragmentation Buffer. Using the mRNA as a template, the first cDNA strand was synthesized with random hexamers, then buffer, dNTPs, RNase H and DNA polymerase I were added to synthesize the second cDNA strand. After that, the cDNA was purified with AMPure XP beads. The purified double-stranded cDNA was then subjected to end repair, A-tailed and connected to sequencing adapters, using the AMPure XP beads for selection of fragment size. Finally, PCR enrichment was conducted to obtain a cDNA library. After the library is constructed, quantitative Real-Time PCR (qRT-PCR) method was used to accurately quantify the effective concentration of the library (library effective concentration > 2 nM) to ensure the quality of the library.