GSM3931375: IgG ChIPSeq; Mus musculus; ChIP-Seq
To prepare chromatin, mouse thymus was extracted from 4-6-week old mice, washed twice with cold PBS and mechanically disaggregated into single cells in 1ml EDTA buffer (100 mM NaCl, 1 mM KH2PO4, 3 mM KCl, 10 mM Na2HPO4, 1 mM EDTA). Thymocytes were then washed twice and resuspended again in EDTA buffer. Cells were fixed by adding formaldehyde to a final concentration of 1% and incubated at 37ºC for 10 min. Libraries were prepared by ChIPmentation as described by Schmidl et al. 2015 using NEBNext HF Polymerase and sequenced using Illumina NextSeq 500.