GSM5470625: dCasMINI-VPR_sgTet_Rep2; Homo sapiens; RNA-Seq
Total RNA was isolated using RNeasy Plus Mini Kit (Qiagen) After the QC procedures, mRNA from eukaryotic organisms is extracted using oligo(dT) beads. Then, the mRNA is fragmented randomly by adding fragmentation buffer, followed by cDNA synthesis with random hexamer primers, a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. After a series of end repair, A tailing and sequencing adaptor ligation, the double-stranded library is completed through size selection and PCR enrichment.