GSM6615076: pool1 Squalene samples; Sus scrofa; RNA-Seq
RNA was extracted from frozen livers   using  Quick-RNA MiniPrep kit (Cat.No:R1055, ZYMO Research, Irvine, CA, USA). Quality control of total RNA sample is assessed by RNA concentration, RIN value,28S/18S and fragment length distribution using Bio analyzer (Agilent RNA 6000 Nano Kit). mRNA enrichment with poly A tail using oligo dT beads. RNA fragmentation and reverse transcription, the first-strand cDNA is generated using random N6-primed reverse transcription, followed by the second-strand cDNA synthesis. Synthesized cDNA is subjected to end-repair and then is 3' adenylated. Adaptors are ligated to the ends of these 3' adenylated cDNA fragments. Followed by several rounds of PCR amplification to enrich the purified cDNA template using PCR primer. Cyclization by denaturing PCR product then DNA is cyclized by splint oligo and DNA ligase. DNA nanoball synthesis and finally, Sequencing on DNBSEQ platform.