2 protocols
Cells were grown in Yeast Extract medium at 30C and harvested from liquid cultures at mid-exponential phase (OD = 0.5) before or after exposure to H2O2.
Cells were crosslinked in 1% formaldehyde for 30 min at room temperature in growth medium and frozen for later use. Pellets were washed in H2O, resuspended in FA lysis buffer (50mM HEPES-KOH pH 7.6, 1mM EDTA pH 8, 150mM NaCl, 1% Triton X-100, 0.1% Na-deoxycholate) and lysed with glass beads in a FastPrep cell disruptor. The extracts were sonicated to fragment chromatin to an average size of ~500 bp with a BioruptorĀ® sonicator and cleared by centrifugation.