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June

23-06-2014

Goal: To obtain single colonies of both Wt and mutant B.Subtilis

We received the wt (1A1) and mutant BKE24040 (isovalerate dehydrogenase beta KO), BKE24050 (isovalerate dehydrogenase alpha KO), BKE24080 (Leucine dehydrogenase KO) from BGSC on 23-06-2014. The wt strain (1A1) was on a paper strip. We received 3 strips from BGSC (Bacillus genetic stock centre). Where as the mutant strains arrived on perti-plates.

Procedure

We plated two strips in two different LBA plates. The plates were free from antibiotics. We placed the strips gently on the LBA and added 1 drop of sterile H2O. After few seconds we streaked the paper strip on the LBA and discarded the strip. The LBA plates were incubated at 37° C. We took single colony from the mutant B.subtilis petri-plates and streaked them on LBA+EM petri plates.

Results

Good growth of both the wt and mutant B.subtilis were observed followed by the overnight incubation

24-06-2014

To obtain single colonies of Wt b.Subtilis from 23-06-14 plate

Procedure

we took a single colony and streaked it on a new LBA with and w/o Erythromycin.

Results

No colonies in LBA + EM. Colonies observed in LBA w/o EM

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October

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