title: GSM5104973: 16_D5_Th1_WT2_Runx1; Mus musculus; ChIP-Seq
library_construction_protocol: Cells (5-20 x 106) were cross-linked for 10 min with 1% formaldehyde, quenched with Glycine, and washed. Cell pellets were resuspended in lysis buffer, and homogenized. Nuclei were resuspended in RIPA buffer and sonicated to a size range of 200-600 bp using Qsonica. Cells were pelleted, and RIPA buffer was added to supernatant to 1 ml. Appropriate antibody coupled to Dynabeads M-280 Sheep anti-rabbit IgG  was added to the mix. The immunocomplexes were washed 5X with LiCl solution.  Supernatants were collected, incubated at 65C overnight, and immunoprecipiated DNA were purified using MinElute columns (Qiagen) Immunoprecipiated DNA was used to prepare library using NEBNext Ultra II DNA library prep kit and NEBNext Multiplex Oligos for Illumina (NEB). ChIP-seq libraries were sequenced with single-end 50 bp using Illumina HiSeq 2500.