GSM5693564: HepG2_RPUSD4KD_mRNA_Rep1; Homo sapiens; OTHER
HepG2 cells were biochemically fractionated and chromatin-associated RNA was isolated and Pseudo-seq was performed. Total RNA was isolated from PUS1 knockout, PUS7 knockdown, RPUSD4 knockdown and wildtype HepG2 cells with Qiazol and poly(A)+ mRNA-seq was perfomed with TruSeq Stranded mRNA kit.   Oligo pools (human, Twist Biosciences) were PCR amplified, and in vitro transcribed into RNA, which was then gel purified, folded, and pseudouridylated with a pseudouridine synthase activity source. Pseudo-seq samples were treated with 0.4 M CMC, followed by reversal of CMC modification in bicarbonate buffer. RT was performed, and truncated cDNAs were size selected, followed by circularization & PCR amplification.