title: KO_AngII1
design: After the total RNA was qualified, the Oligo(dT) -linked magnetic beads were used to enrich eukaryotic mRNA. The extracted mRNA was randomly broken into short fragments by Fragmentation Buffer. The fragments of mRNA were used as template, and a strand of cDNA was synthesized by Random hexamers. Buffer, dNTPs, RNaseH, and DNA Polymerase I were subsequently added for two-strand cDNA synthesis. AMPureXP beads were used to purify the double-stranded product, T4 DNA polymerase and Klenow DNA polymerase activities were used to repair the sticky end of the DNA to A flat end, base A was added to the 3 'end and an adapter was added, AMPureXP beads were used for fragment selection, and T4 DNA polymerase and Klenow DNA polymerase activities were used to repair the sticky end of the DNA to a flat end. Finally, PCR amplification was performed to obtain the final sequencing library. After qualified quality inspection, the library was sequenced by Illumina Novaseq 6000 with a double-end 2*150bp (PE150) read length.