RNA-Seq of piPSCs
Total RNA was treated using Oligo dT-enriched mRNA and purification. Fragmented RNA was then reverse-transcribed using random N6 primers to form double-stranded DNA (second-strand cDNA synthesis with dUTP instead of dTTP). Next, 3'adenylated and adaptor ligation was added to the end of the synthesized double-stranded DNA, which was amplified using PCR with specific primers. The PCR products were thermally denatured into a single strand and a bridging primer was used to form a loop to obtain a single-stranded circular DNA library