title: GSM1916156: C2C12_ZFP932_input_R2; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were harvested, washed with Episerf, fixed in 2mL per 1x107 cells (10 min in 1% formaldehyde), quenched with glycine in 10mL (at 125 mM final), washed three times with PBS, and pelleted. Each pellet containing 1x107 cells was lysed, resuspended in 1 mL of sonication buffer on ice (10 mM Tris at pH 8, 1 mM EDTA, 0.1% SDS, and protease inhibitors), transferred to TC 12x12 tubes (Covaris), and sonicated (Covaris settings: 20 min, 5% duty cycle, 140W, 200 cycles). Sonication was assessed by reverse cross-linking (65oC, RNAse A at 1µg/µL, overnight), followed by DNA extraction. Fragment size (between 200-400bp) was checked on a Bioanalyzer (Agilent 2100). Immunoprecipitations were performed with chromatin from 4x107, with Dynabeads (Protein G, ThermoFisher) in IP buffer (10 mM Tris at pH 8, 1 mM EDTA, 0.1% SDS, 150 mM NaCl, 10% Triton X-100, and protease inhibitors) overnight. Chromatin was reversed cross-linked (65oC, Proteinase K at 400ng/µL, overnight) and DNA was extracted. Single-end libraries were prepared and sequenced in 100bp reads runs according to Illumina instructions.
attributes.tag: GEO Accession
attributes.value: GSM1916156