GSM7814149: iCAF_1; Homo sapiens; RNA-Seq
Total RNA was extracted using Trizol (Thermo Fischer) and a spin column method according to the manufacturer's instructions (Zymo Research) DNAse treated total RNA samples were sent to BGI, Hong Kong for RNA-seq. There, total RNA was enriched for mRNAs using oligo dT magnetic beads. The mRNAs were fragmented and reverse transcribed to double-strand cDNA by random-hexamer primers. The synthesized cDNA was subjected to end-repair and was then 3'Adenylated. Adaptors were ligated to the ends of these 3' Adenylated cDNA fragments. The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer, splint oligo and DNA ligase, followed by sequencing on the DNBseq platform resulting in on average ~30 million reads per sample.