GSM8796515: Human kidney, normothermic perfusion, RBC, 48 hrs, HF20, donor 4; Homo sapiens; RNA-Seq
RNA was isolated using the QIAGEN mini RNA isolation kit according to manufacturer's protocol, subsequently mRNA was enriched for poly-A tails using oligo(dT)-magnetic beads. After enrichment, mRNA was fragmented using fragmentation buffer, followed by reverse transcription using random N5 primers to synthesize cDNA double strands. The synthesized double-stranded DNA was then end-repaired and 5' phosphorylated, with a protruding 'A' at the 3'end forming a blunt end, followed by ligation of a  bubble-shaped adapter with a protruding 'T' at the 3' end. The ligation producs were PCR amplified using specific primers. The PCR products were denatrued to single strands, and then single-stranded circular DNA libraries were generated using a bridged primer. The constructed libraries were quality-checked and seuenced after passing the quality control. (BGI's DNB-seq protocol)