DNA from DnaA-pulldown of HPUra-treated cels
The protocol for immunoprecipitation of His-tagged DnaA was largely based on Minnen et al., Mol. Micro, 2011. Specifically, cells were pre-cultured in acid C+Y (pH 6.8) and grown until OD600 0.40. Cells were then diluted 1:50 in acid C+Y, to a final volume of 200 mL and grown until OD600 0.12. The culture was then split into two, 0.15 ug/mL HPUra was added to one half and an equivalent volume of 10 mM NaOH to the other. Fixation, formaldehyde quenching and harvesting was performed as described earlier.