GSM2771537: K562 ZBTB7A ChIP-seq Input 1; Homo sapiens; ChIP-Seq
ChIP experiments were performed as previously described (Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. ChIP-seq: using highthroughput  sequencing to discover protein-DNA interactions. Methods. Jul 2009;48(3):240-248). Briefly, approximately 5x107-1x108 cells were used for each IP. Cells were crosslinked with 1 % formaldehyde for 10-15 min at RT and the reaction was quenched with glycine at a final concentration of 125 mM. Crosslinked cells were then lysed and sonicated to obtain ~200–300 bp fragments of chromatin. Crosslinked DNA was pulled down at 4 °C overnight using antibody against V5-tag (R960CUS, Life Technologies), ZBTB7A (Santa Cruz Biotechnology Inc, sc-33683 x and eBioscience, 14-3309-82) or control anti-goat IgG (sc-2028). Chromatin was then reverse crosslinked and eluted at 65°C overnight and DNA was purified.If ChIP material was subjected to sequencing, DNA was purified using a Qiagen MinElute Reaction Clean-up kit. Standard Illumina TruSeq