title: GSM6721011: Trim33WT_HU_rel; Mus musculus; OTHER
library_construction_protocol: The assay was performed as described previously (Lensing et al, 2016) with modifications. Cells were fixed with formaldehyde and permeabilized with 0.2% Triton X-100 in PBS. DNA end repair reaction was performed by incubation with E.Coli DNA Polymerase I Klenow fragment for 30 minutes at 25C. Cells were washed and A-tailing reaction was performed in dA-tailing buffer (NEB) with Klenow exo- for 30 minutes at 37C. dA-tailed DNA fragments were ligated to a biotinylated NEB adapter at 16C overnight. Cells were lysed in 20mM Tris pH8.5, 300mM NaCl, 0.2%SDS and treated with proteinase K, followed by sonication, phenol-chloroform extraction and ethanol purification of DNA. Biotinylated DNA fragments were captured on Streptavidin beads and used for library preparation with NEB Ultra II DNA library end prep kit. Sequencing libraries were prepared using NEB Ultra II DNA library Prep kit according to the manufacturer's instructions. DSB capture and sequencing