RNA-Seq analysis between AAV-High and AAV-Cre hearts

Oligo dT microbeads were used to enrich mRNA with poly A tail. The resulting mRNA was randomly interrupted and reverse-transcribed by a random N6 primer to generate a first-strand cDNA, and then a second-strand cDNA is synthesized with dUTP instead of dTTP. The synthesized cDNA was end-repaired and then 3' adenylation was performed. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments. The dUTP-marked strands were selectively degraded by Uracil-DNA-Glycosylase (UDG). The remaining strands were amplified to generate a cDNA library suitable for sequencing. The library was sequenced on a DNBSEQ (DNBSEQ Technology) platform.