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On the structured plates, this was achieved after replica plating from viable counts of the remaining lawn; colonies were differentiated by colony morphology and pigmentation.
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All steps involving bacterial transformations used MegaX DH10B competent cells (Life Technologies) and yielded lawns of colonies (>50 000 per transformation) whose numbers were always >10-fold larger than the number of variants in each library.
The brown halos appearing around some of the filters are due to toxicity of the compounds deposited onto these filters which, by preventing yeast cell growth and therefore appearance of a lawn of yeast colonies (either as white or red colonies) reveals the brown/orange colour of the YPD medium.
A 10 µL loop of bacteria from the lawn of grown colonies was suspended in 500 µL of TE buffer and incubated at 95°C for a further 10 min.
Absence of growth of the bacterial lawn around a colony results in an inhibition zone.
Hyphae form uniform lawn over the white colony, mycelia aerial comprising short hyphae in the form of a uniform lawn over the colony.
Above this lawn, the earliest Lac+ revertant colonies appear on day 3.
Nontoxic concentrations were categorized as those where there was a well-developed lawn, almost similar size of colonies and no statistical difference in the number of spontaneous revertants in test and control plates.
Non-toxic concentrations were categorized as those where there was a well-developed lawn, almost similar size of colonies, and no statistical difference in the number of spontaneous revertants in test and control plates.
To avoid the disturbance of mineral medium on biofilm morphology (dome-shaped colonies surrounded by lawns like cells), biofilms were grown under the condition without biomineralization medium.
Further, tpp1− cells were less viable after culture in ASM, as assayed by the ability to form colonies on bacterial lawns (Fig. 6B).
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