MethylC-seq or BS-seq

Cited from the paper "MethylC-seq library preparation for base-resolution whole-genome bisulfite sequencing"

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MethylC-seq library preparation protocol overview. gDNA (i) is fragmented to ~200 bp by sonication (ii). DNA fragments containing damaged or incompatible 5′- and/or 3′-protruding ends are converted to 5′-phosphorylated, blunt-ended DNA (iii). Blunt-ended DNA fragments are converted to DNA with 3′-dAMP overhangs (iv). Methylated Y-shaped adapters are ligated to the dA-tailed DNA fragments (v). All cytosines in the adapters must be methylated to allow for primer binding and amplification after bisulfite conversion. Adapter-ligated DNA fragments are denatured, and unmethylated cytosine is converted to uracil during sodium bisulfite treatment (vi). Bisulfite-treated DNA fragments remain single-stranded as they are no longer complementary. Low-cycle PCR amplification is performed with a polymerase that can tolerate uracil residues (vii). The final library fragments contain thymines and cytosines in place of the original unmethylated cytosines and methylated cytosines, respectively (viii).
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