GSM3895484: ChIP-seq of Iss1-GFP from clr4∆ cells; Schizosaccharomyces pombe; ChIP-Seq
Cells were fixed with 1% formaldehyde at room temperature for 20 minutes. Additional crosslinking with DMA were performed as needed for in each sample as indicated in each description section. Cells were lysed using bead-beater method. Lysates were sonicated and immunoprecipitations were performed in cold room conditions. Whole-cell extracts (WCE) were used as input control for each ChIP experiment. Reverse crosslinking was performed overnight at 65˚C, followed by treatment with RNase A and proteinase K. DNA was purified using QIAquick PCR purification kit (cat. no. 28014). NEBNext Ultra II DNA Library Prep kit (NEB, E7645S) was used according to manufacturer's recommendation. Libraries were analyzed using an Agilent 2100 BioAnalyzer (Agilent) and sequenced on the Illumina MiSeq or NextSeq 500 platform.