GSM2544917: hos1_aid_10; Saccharomyces cerevisiae; ChIP-Seq
For crosslinking, cells were incubated with 1% formaldehyde overnight at 4°C. Cells were harvested and washed three times with ice-cold TBS, and cells were transferred to 1.5 ml tube. For cell breakage, 300 ml of lysis buffer and glass beads were added to the tube, and placed tubes into the cells disruptor (FastPrep-24 5G), and disrupt cell wall at speed 6.0 m/sec for 40 sec, repeat the disruption procedure once more to achieve better results. The cell lysate was sonicated with Bioruptor (Diagenode, 30sec ´ 30 cycles with 30 sec intervals with high intensity), resulting in an average fragment size of 300~200 bp. The lysate was clarified by centrifugation at 13,000 rpm for 5 minutes. For immunoprecipitation, 60 ml of magnetic beads (Dynabeads Pan Mouse IgG, Invitrogen) were added following pre-incubatition with an anti-HA monoclonal antibody (HA.11, Covance).