N2-heptynyl-dG sequencing

N2-heptynyl-dG sequencing in HEK293T cells HEK293T cells were treated with N2-heptynyl-dG for 3-h at a final concentration of 10 μM. The cells were then harvested immediately for N2-heptynyl-dG sequencing. The genomic DNA were extracted using Monarch® Genomic DNA Purification Kit (New England Biolabs) and reacted with biotin-azide at room temperature for 6 h. The biotinylated genomic DNA was sheared using a Covaris S220 sonicator. The N2-heptynyl-dG DNA were enriched with Dynabeads™ MyOne™ Streptavidin C1 (Thermo). Purified DNA was quantified and verified on an Agilent 2100 Bioanalyzer. The library was then constructed using NEBNext Ultra DNA Library Prep Kit (NEB, E7103S) following the manufacturer’s instructions. Subsequently, the purified DNA libraries were assessed using an Agilent 2100 Bioanalyzer and multiplexed for sequencing on a DNBSEQ-G400 (BGI Genomics) .