GSM6935305: DMSO_Input_3_Rep1; Saccharomyces cerevisiae; ChIP-Seq
ChIP was performed essentially as described previously (70,71). Briefly, 200 or 250 mL of cell cultures were grown to log phase (OD600 = 0.8 - 1.0). Cells were crosslinked for 20 min at room temperature by addition of formaldehyde to 1% (v/v). Lysates were prepared in FA lysis buffer (72). Chromatin was sheared in Bioruptor® Pico 15 mL tubes with 300 µL beads (Diagenode C30010017) by 25 cycles of 30 sec on and 30 sec off using a Bioruptor® (Diagenode B01060010). Sonicated chromatin was brought up to a volume of 6 mL in FA lysis buffer and centrifuged at 50,000 rpm for 20 min at 4˚C. Chromatin in the supernatant was aliquoted, flash-frozen, and stored at -80˚C. S. cerevisiae chromatin was mixed with Schizosaccharomyces pombe chromatin (KP07 or KP08) at a 1:10 ratio (S. pombe : S. cerevisiae) based on protein concentration (Pierce BCA protein assay kit; Thermo Fisher Scientific 23227), and IP reactions were conducted with anti-V5 (Invitrogen R960-25, 1 µL/225 µg or 10 µL/1800 µ