GSM6938440: TAPSbeta_d8DZPGCLC_2; Homo sapiens; Bisulfite-Seq
Genomic DNA was purified using Quick-DNA mini prep kit (ZYMO research). DNA was spiked with spike-in control DNA and sonicated to 300-500 bp, before ligated with NEBNext Adaptor for Illumina using KAPA HyperPrep Kit according to the manufacturer's protocol. The uracil in the loop of NEBNext Adaptor was removed by USER Enzyme (New England Biolabs). 100 ng ligated DNA was used for both TAPSβ and CAPS. For TAPSβ, the ligated library was subjected to βGT (Thermo Fisher) blocking, two rounds of mTet1 oxidation, and borane reduction. For CAPS, the ligated library was subjected to chemical oxidation and borane reduction. Converted DNA from TAPSβ and CAPS was amplified with NEBNext Multiplex Oligos for Illumina and KAPA HiFi HotStart Uracil+ ReadyMix PCR Kit for 4 cycles according to the manufacturer's protocol. The PCR product was purified with Ampure XP beads and quantified with Qubit dsDNA HS Assay Kit (Thermo Fisher) according to the manufacturer's protocol. Libraries were sequenced for 150 cycles in paired-end mode on the NovaSeq platform.