Name: GSM6052747
Instrument: Illumina HiSeq 4000
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: SINGLE
Construction protocol: For LAMS-HTGTS, genomic DNA was isolated from WT, Setx-/- , Rnaseh2bf/f , and Setx-/- Rnaseh2bf/f cells 72 hours after stimulation to switch to IgG1 with 72 h LPS/IL-4/α-RP105. For DRIP-seq, genomic DNA was gently extracted by SDS/Proteinase K treatment at 37°C followed by phenol-chloroform extraction and ethanol precipitation. The extracted genomic DNA was fragmented with restriction enzyme (Hindlll, Xbal, EcoRI, SspI, BrsGI), 4 micrograms of digested DNA were incubated with 2 micrograms of S9.6 antibody overnight at 4 ℃ in DRIP buffer (10 mM NaPO4 pH 7.0, 140 mM NaCl, 0.05% Triton X-100). In vitro RNase H digestion was used to generate a negative control. After incubation, antibody-DNA complexes were bound to Protein G Dynabeads and thoroughly washed, DNA was recovered with Chelex-100, the bound fraction was suspended in 0.1 ml 10% Chelex-100 (Bio-Rad), vortexed, boiled for 10 min, and cooled to room temperature. This sample was added to 4 μl of 20 mg/ml Proteinase K followed by incubation at 55°C for 30 min while shaking. Beads were boiled for another 10 min. Sample was centrifuged and supernatant collected. Beads were suspended with 100 μl 2× TE to the beads, vortexed, centrifuged, and supernatants pooled. For LAMS-HTGTS, linear amplification-mediated high-throughput genome-wide translocation sequencing (LAM-HTGTS) libraries were prepared from genomic DNA as previously described (Hu et al., 2016; Yin, Liu, Liu, & Hu, 2019; Yin, Liu, Liu, Wu, et al., 2019). For DRIP-seq, libraries were prepared as described in (Ginno et al., 2012; Sanz et al., 2016)