title: RNA-Seq of mus musculus: adult female bone marrow
design: Total RNA was extracted using the TRIzol reagent according to the manufacturer's protocol. mRNA enrichment was performed by using OligodT magnetic beads to enrich polyA-tailed mRNA. The obtained RNA was fragmented using a fragmentation buffer, and then reverse transcription was carried out with random N6 primers to synthesize the first-strand cDNA. Subsequently, the second-strand cDNA was synthesized to form double-stranded DNA. The synthesized double-stranded DNA was end-repaired and phosphorylated at the 5' end, with a protruding 'A' base formed at the 3' end. A bubble-shaped adapter with a protruding 'T' base at the 12' end was then ligated. The ligation product was amplified by PCR using specific primers. The PCR product was thermally denatured into single-stranded DNA, and then a bridge primer was used to circularize the single-stranded DNA to obtain a single-stranded circular DNA library. After quality inspection of the constructed library, sequencing was performed if the quality met the requirements.