GSM8615110: Wild type Dnmt1 rep3; Mus musculus; OTHER
Genomic DNA was purified from cell lysates using Zymo Genomic DNA Clean & Concentrator-10 Kit. Libraries were constructed according to the TOP-seq (Tethered Oligonucleotide-Primed sequencing) protocol (Stasevskij et al., 2017). Briefly, 900 ng of sonicated genomic DNA were end-filled, a 3'-dA mononucleotide extension was added and DNA was ligated to adapters. Ligated DNA was then subjected to Cu(I)-catalyzed cycloaddition reaction in which the alkyne-bearing oligodeoxyribonucleotide with biotin was attached to the azide moiety of the modified DNA. Biotinylated DNA was enriched with streptavidin-coupled magnetic beads and subsequently used in a TOP DNA priming reaction from the attached priming oligodeoxyribonucleotide. The products were amplified and subjected to Ion Proton sequencing.