DNA methylation of Odorrana tormota: G45 male hindlimb thigh muscle
After the sample is qualified, the DNA sample is first digested with methylation-insensitive restriction enzyme, MspI; the end of the digested DNA fragment is repaired, A tail is added, and all cytosine-modified sequencing adapters are connected. ; Cut the gel to select a DNA fragment with an insert length ranging from 150-300bp; then perform Bisulfite treatment (using EZDNAMethylationGoldKit, ZymoResearch). After processing, the unmethylated C becomes U (the PCR becomes T), while the methylated C remains unchanged. Finally, PCR amplification is performed to obtain the final DNA library.