title: ChIPseq of mouse: Min6 cell line input control
design: 10 mio Min6 cells were used per ChIP. Cells were crosslinked with 1 % formaldehyde for 10 min at room temperature, crosslinking was stopped by addition of 2M glycine (0.125 M final concentration) at room temperature for 5 min. Cells are washed twice in cold 1x PBS. Cells were lysed into ice-cold lysis buffer for 1h on ice. Following cell lysis, the samples were sonicated to generate fragments of average length of 300-100 bp. Cellular debris was removed by, 10 % of the lysate was stored as the source of the input sample and the remaining lysate was diluted, in the presence of 70 microliter of anti-v5 magnetic beads were then incubated at 4 degree C with rotating overnight. Beads were washed extensively at 4 degree C in low salt buffer, in high salt buffer, and in LiCl buffer, and finally in 1 ml of TE buffer. The bound chromatin was released from the beads by vortexing at room temperature in 200 ?l elution. RnaseA and NaCl (200mM final concentration) was added to the eluate and followed by reverse crosslinking by heating at 65 degree C o/N and proteinase K digestion. DNA was purified using a QIAGEN PCR purification kit.