Transcriptome sequencing of muscle
Total RNA was extracted from a sample of muscle tissue at the back of the rabbit's neck using the Trizol method, and RNA purity was tested with a Nano Photometer spectrophotometer (IMPLEN, Westlake Village, USA). Construct cDNA libraries from 1 g of total RNA using the cDNA-PCR sequencing kit (SQKPCS109). Reverse transcriptase is used to enrich full-length cDNA and add defined PCR linkers at both ends of the first strand of cDNA, followed by 14 cycles of cDNA PCR with LongAmp Tag DNA polymerase (New England Biolabs, Ipswich, MA, USA) (extension time 8 min). The PCR products were then ligated using T4 DNA ligase (New England Biolabs, Ipswich, MA, USA) for ONT linkage. Agencourt XP beads (Beckman Coulter, Brea, USA) are used for DNA purification. The final cDNA library was processed with FLOMIN109 flow cells and analyzed on Biomarker Technology Company's PromethION platform in Beijing, China.