Synchronization of Bacillus subtilis Cells by Spore Germination and Outgrowth

This protocol defines conditions under which the germination of spores can be used to synchronize Bacillus subtilis cells, utilizing the time-ordered sequence of events taking place during the transition from spore to vegetative cells. The transition stages involve: phase change, swelling, emergence, initial division, and elongation. By using this method we have obtained two distinctive synchronized cell cycles, while the synchrony faded away in the third cycle. The advantage of using spore outgrowth and germination is that a highly synchronized population of bacterial cells can be obtained. Non-dividing spores stay synchronized, while synchrony rapidly decays during a few divisions. The limitations of this method are that it can be applied only for sporulating bacteria and synchrony lasts for only a limited period of time exceeding not more than two cycles.