GSM2576786: chip1_wce_rct1_2; Schizosaccharomyces pombe; ChIP-Seq
Cells were spheroplasted by zymolyase at 37°C and then sonicated using a bioruptor for 8 cycles (30s ON 60s OFF). For each IP, 500-750 ug of chromatin were used with 3 to 5 ul antibody. 1 ng of DNA purified from ChIP experiments was made into libraries by using NEB enzymes. In brief, DNA was end-repaired by T4 DNA polymerase, Klenow fragment and T4 DNA PNK. “A” bases were added to the 3’ end of end-repaired DNA fragment with Klenow 3’ to 5’ exo minus and dATP. Barcoded Truseq adaptors (Illumina) were ligated to DNA fragments using quick ligase at 25°C. Five PCR cycles were performed prior to size selection. After size selection, purified DNA was PCR amplified with 6 to 12 cycles (Kapa HiFi HotStart ready mix). Barcoded libraries were pooled and sequenced on Illumina HiSeq platform.