GSM8889320: CON5 (L5); Bos taurus; RNA-Seq
According to the manufacturer`s instructions, total RNA was obtained from liver tissue using TRIzol reagent (Molecular Research Center, Cincinnati, Ohio, USA) RNA reverse transcription, library preparation, and RNA sequencing (RNA-seq) were carried out at BGI Co., Ltd. (Shenzhen, China). Briefly, total RNA was enriched for mRNA by poly(A) selection using oligo(dT) magnetic beads, followed by reverse transcription and cDNA synthesis. The resulting double-stranded cDNA underwent end repair, 5'-phosphorylation, and 3'-adenylation to prepare for adapter ligation. The adapter-ligated products were then PCR amplified, denatured, and circularized using bridging primers to create single-stranded circular DNA libraries. Sequencing was performed on the DNBSEQ platform with PE500 (BGI Co., Ltd., Shenzhen, China). The raw sequencing data were processed using SOAPnuke (v1.5.6, RRID:SCR_015025) to eliminate low-quality and adapter-contaminated reads. The filtering criteria were as follows: 1) removal of reads containing adapter sequences, 2) exclusion of reads with more than 5% unknown bases (N), and 3) discarding low-quality reads, defined as those in which more than 20% of the bases had a quality score below 15.