GSM7325155: DKO, 12h, RPF; Homo sapiens; RNA-Seq
The Ribosome protected fragment (RPF) was isolated by 34% sucrose cushion and UREA-TBE gel size selection. Total RNA and RPF were then extracted using TRIzol regent. Both fragmented total RNA and ribosome-protected RNA fragment were subjected to library preparation following standard procedure of ARTseq™ Ribosome Profiling Kit with minor modification. In brief, RNA samples were end repaired with T4 PNK and ligated with 3' adaptor, followed by cDNA synthesis and cDNA purification. The cDNA was then subjected to circularization and PCR amplification.