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Plates were incubated overnight at 37°C and the resulting colony forming units (CFU) were counted.
After an incubation period of up to 72 h (37°C), resulting colony forming units (CFU) were counted, subcultivated, and identified according to standard microbiological procedures (API20E-systems, Fa. API Biomerieux SA, Marcy l'Etoile, France).
If an intramolecular integration event occurs, the lacZ gene located between the attB and attP sites will be excised, and a resulting colony will be white.
Deletion of any of the four genes therefore results in QS-deficient mutants that behave as obligate cheaters that are unable to fully co-operate, the resulting colony morphologies are shown in the diagonal of Figure 1B.
The resulting colony filters were fixed by treatment with a series of four solutions in the following manner.
A sample was streaked to Marine-Fe Agar, incubated for 2 d, and a resulting colony of a spiral bacterium (termed strain HJ) was subcultured into Marine-Fe broth.
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After incubating overnight, resulting colonies were screened by PCR.
The resulting colonies were isolated and purified on Zobell marine agar medium using serial dilution technique.
Rescreening the colonies for a single crossover on a new plate supplemented with kanamycin showed the double cross over pattern for most of the resulting colonies.
When pig fecal samples were plated on KMI agar medium, approximately 60 100% of the resulting colonies tested positive using the M. elsdenii-specific PCR primers.
After the 1 month infection, all resulting colonies were lysogenic and assembled into biofilms.
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