Paris/July 6
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== Preparing growth medium for transduction screening == | == Preparing growth medium for transduction screening == | ||
If the transduction works (i.e. an homologous recombinaison that should delete DapA by inserting an erythromycin resistant cassette), transducted MG1655 Coli should grow in a medium LB+erythro+ DAP but can't grow in a medium LB+erythro without DAP. | If the transduction works (i.e. an homologous recombinaison that should delete DapA by inserting an erythromycin resistant cassette), transducted MG1655 Coli should grow in a medium LB+erythro+ DAP but can't grow in a medium LB+erythro without DAP. | ||
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Nile Red can be excited by light around 312nm (Spiekermann,1999). | Nile Red can be excited by light around 312nm (Spiekermann,1999). | ||
| - | After 24h incubation, we observed Acinetobacter and E. | + | After 24h incubation, we observed Acinetobacter and E.coli on LNMM with and without Nile Red: |
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| + | [[Image: Acineto_Coli_24h.jpg|center|300px]] | ||
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| + | We can observe basic fluorescence with Acinetobacter on LNMM with and without Nile Red. With E.coli transformed or not with pKS::DGAT, we do not observe fluorescence; indeed, coli do not grow on LNMM... | ||
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| + | We decided to wait more to see acccumulation of triglyceride with Acinetobacter. | ||
[[Paris|<<home]] | [[Paris|<<home]] | ||
Latest revision as of 19:00, 3 October 2007
Preparing growth medium for transduction screening
If the transduction works (i.e. an homologous recombinaison that should delete DapA by inserting an erythromycin resistant cassette), transducted MG1655 Coli should grow in a medium LB+erythro+ DAP but can't grow in a medium LB+erythro without DAP.
- Preparation of DAP solution from the powder (50mM). See Protocols
- Making 10 petri dish (LB+tet+citrate+DAP). See Protocols
- Making 10 petri dish (LB+erythromycin+citrate+DAP). See Protocols
Acinetobacter and E.Coli on LNMM Nile Red solid culture
Nile Red can be excited by light around 312nm (Spiekermann,1999). After 24h incubation, we observed Acinetobacter and E.coli on LNMM with and without Nile Red:
We can observe basic fluorescence with Acinetobacter on LNMM with and without Nile Red. With E.coli transformed or not with pKS::DGAT, we do not observe fluorescence; indeed, coli do not grow on LNMM...
We decided to wait more to see acccumulation of triglyceride with Acinetobacter.
