GSM5171132: Input 2; Homo sapiens; ChIP-Seq
2x10^7 cell were crosslinked with 1% formaldehyde in CiA fixation buffer (50mM HEPES pH8, 1mM EDTA, 0.5mM EGTA, 100mM NaCl) for 10min. Then glycine was added to a final concentration of 125mM to stop the fixation. To collect nuclei cells were resuspended in CiA NP-Rinse 1 (50mM HEPES pH8, 140mM NaCl, 1mM EDTA, 10% glycerol, 0.5% NP40, 0.25% Triton X-10) and incubated in ice for 10min. Cells were washed with CiA NP-Rinse 2 (10mM Tris pH8, 1mM EDTA, 0.5mM EDTA, 200mM NaCl) and dissolved in Covaris shearing buffer (0.1% SDS, 1mM EDTA, 10mM Tris-HCl pH8). Cells were sonicated using Covaris S220 (Intensity 4, Duty Factor 5%, Cycle/Burst 200) for 420sec. Libraries were prepared using NEBNext Ultra II DNA Library Prep kit for Illumina (NEB #E7103) and NEBNext Multiplex Oligos for Illumina (NEB #E7730S). To control the quality of DNA libraries, the size and purity of samples were confirmed by Bioanalyzer.
GEO Accession
GSM5171132