GSM7990613: RNAseq_CON_B_#1; Bos indicus x Bos taurus; RNA-Seq
Total RNA was isolated from bovine fibroblast cells using TRIzol™ Reagent (Invitrogen, 15596026) following the manufacturer's instructions. The concentration of RNA was measured by using a NanoDrop® ND-1000 Spectrophotometer. RNA samples were stored at -80°C. Total RNA-seq for fibroblast cells was conducted by CD Genomics. Information on library preparation and sequencing obtained from the company is as follows: The first step involves the removal of ribosomal RNA using RiboZero kit (Illumina). Subsequently, the RNA is fragmented into small pieces using divalent cations under elevated temperatures. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis. These cDNA fragments then have the addition of a single 'A' base and subsequent ligation of the adapter. The products are purified and enriched with PCR to create the final cDNA library. This results in libraries with inserts ranging in size from 120–200 bp with a median size of 150 bp. Equimolar pooling of libraries was performed based on QC values and sequenced on Novaseq6000 sequencer (Illumina) with a read length configuration of 150 PE for 33M PE reads per sample (33M in each direction).