Glycerol) and rotated for 30 min at area temperature. Each mixture was transferred to a Poly-Prep column (Bio-Rad, Hercules, CA), and washed 3 times with 1 ml equilibration/wash buffer, with care taken to ensure that all washes had been done below virtually identical conditions. Complexes of DnaA-his bound to DNA had been eluted by adding 0.five ml ChIP elution buffer (50 mM Tris-HCl, pH 8.0, ten