title: GSM6103026: Tet3 Mut 1; Mus musculus; OTHER
library_construction_protocol: Genomic DNA extracted from cells were eluted in nuclease-free water. Illumina sequencing library was generated from 75 ng of genomic DNA using the NxSeq UltraLow DNA Library Kit (Lucigen) following the manufacturer's instruction, but without PCR amplification. Glucosylation of half of the purified DNA library was performed in 21 µL reaction containing 1× Thermo Epi Buffer, 100 µM N3-UDP-Glc and 2 µL of T4 beta-glucosyltransferase (Thermo Fisher) at 37 °C for 1 hr. After glucosylation, 2.1 µl of Biotin-PEG4-DBCO (Click Chemistry Tools, 20 mM stock) was added directly to the reaction mixture, and the mixture was incubated at 37 ˚C for 2 hr. The biotinylated DNA was purified by (1.8× volume) paramagnetic DNA binding beads (Omega Bio-Tek) according to standard procedure. The purified DNA was incubated with 5 µl of C1 Streptavidin beads (Life Technologies) in B&W buffer (B&W buffer: 5 mM Tris [pH 7.5], 0.5 mM EDTA, and 1 M NaCl) according to the manufacture's instruction, for 40 min at room temperature with rotation. The beads were subsequently undergone six 5-min washes with B&W buffer before finally eluted in 40 µL water. The eluted enriched DNA libraries were PCR amplified with index primers and sequenced on an Illumina HiSeq 2000.