GSM8988487: cKO1; Mus musculus; RNA-Seq
RNA was extracted from tissues using standard extraction methods, and RNA samples were subjected to strict quality control by Agilent 2100 bioanalyzer to precisely detect RNA integrity. The first strand cDNA was synthesized in M-MuLV reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primers, followed by RNA strand degradation with RNaseH, and the second strand cDNA was synthesized from dNTPs in DNA polymerase I system. The purified double-stranded cDNA was end-repaired, A-tailed and ligated with sequencing adapters, and the cDNA of about 250 – 300 bp was screened with AMPure XP beads, amplified by PCR and the PCR product was purified again using AMPure XP beads to finally obtain the library.