RNA-Seq of gastrocnemius
Use mRNA enrichment method to process total RNA, mRNA enrichment: Use magnetic beads with OligodT to enrich mRNA with polyA tail. Fragment the obtained RNA with the interrupted buffer, reverse transcription with random N6 primers, and synthesize cDNA strands to form double-stranded DNA. The synthetic double-stranded DNA ends are filled in and the 5'end is phosphorylated to form a sticky end with a protruding "A" on the 3'end, and then a blister-like linker with a protruding "T" on the 3'end is connected. The ligation product is amplified by PCR with specific primers. The PCR product is thermally denatured to single-stranded, and then the single-stranded DNA is circularized with a bridge primer to obtain a single-stranded circular DNA library.
S3-R