GSM2824339: WT rep2; Schizosaccharomyces pombe; ChIP-Seq
The frozen pellet was resuspended in 500 µl lysis buffer: 50 mM Hepes pH 7.4, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% Sodium Deoxycholate, 1% PMSF and EDTA-free protease inhibitor cocktail (Roch) and lysed using FastPrep (MP Biomedical; setting: 5x 30 Sec cycles at 4.5 m/s). Each lysate was recovered in a 5 ml snap-cap tube by centrifugation at 1500 rpm for 3 min and sonicated using the Covaris E210 instrument. Sonication settings used were: 20%, 8 intensity, 200 cycles per burst for 12 minutes. Libraries were prepared using DNA SMART ChIP-Seq Kit (Clontech) and Illumina indexed oligomers. Following PCR amplifications (15 cycles), libraries were size selected and purified using Agencourt AMPure XP beads (200-400 bp range). Each library was validated using Quibit 2.0 Fluorometer for concentration, Agilent 2100 Bioanalyzer for fragment size, and NEBNext Library Quant Kit (New England BioLabs) for adaptor presence. Libraries from 12 samples were pooled in equimolar ratio (10 nM) and submitted for sequencing using Illumina HiSeq2500 Rapid Run mode.
GEO Accession
GSM2824339