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. 2013 Jul 16;8(7):e69491. doi: 10.1371/journal.pone.0069491

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© 2013 Seike et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Wild-type P-cells (FS127), wild-type M-cells (L972) and M-factor-less M-cells (FS120) were mixed in SSL−N in the cell number ratio of 2:1:1. FS127 carries a uracil-auxotrophic marker (ura4-D18), and FS120 harbors an adenine-auxotrophic marker (ade6-M210). After 4 hr of culture with shaking, cell aggregates were obtained after the culture was allowed to stand for a few minutes. Cells were sampled from the precipitated aggregates and the floating fractions separately. The fractions were briefly sonicated to disperse cell clusters, and the cells were spread on SD without leucine but with one-quarter strength adenine-sulfate. P-cells could not form colonies because of uracil depletion. M-cells derived from the M-factor-less strain formed red colonies due to ade6-M210, and those from the wild-type strain formed white colonies.