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Team:Paris Liliane Bettencourt/Notebook/2011/09/02/
From 2011.igem.org
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Electroporation of Danyel's S81 in pHM3 seems to be a success. However, the positive controls grew even on 75 μg.ml<sup>-1</sup> without the pHM3 resistance to TetR. This brings about a BIG problem: our wild strain is resistant to high levels of tetracycline either because the tet(l) gene mutated (name of gene might be incorrect but natural tetR resistance gene exists) or the strain we were given was already very resistant. The other possibility is that it was given with a plasmid :: TetR, the last possibility is that our tetracycline is non-functional (photodegradation, others ?). In any case, I doubled and quadrupled concentrations and replated as little biofilm as possible of each of the transformants. | Electroporation of Danyel's S81 in pHM3 seems to be a success. However, the positive controls grew even on 75 μg.ml<sup>-1</sup> without the pHM3 resistance to TetR. This brings about a BIG problem: our wild strain is resistant to high levels of tetracycline either because the tet(l) gene mutated (name of gene might be incorrect but natural tetR resistance gene exists) or the strain we were given was already very resistant. The other possibility is that it was given with a plasmid :: TetR, the last possibility is that our tetracycline is non-functional (photodegradation, others ?). In any case, I doubled and quadrupled concentrations and replated as little biofilm as possible of each of the transformants. | ||
| - | + | <br> | |
In parallel, I am going to test a simple construct to test efficiency | In parallel, I am going to test a simple construct to test efficiency | ||
| - | pVEG-SpoVG-RFP-terminator | + | pVEG-SpoVG-RFP-terminator <br> |
bacterial strains were inoculated today. | bacterial strains were inoculated today. | ||
Revision as of 17:03, 2 September 2011
Adrien
Electroporation of Danyel's S81 in pHM3 seems to be a success. However, the positive controls grew even on 75 μg.ml-1 without the pHM3 resistance to TetR. This brings about a BIG problem: our wild strain is resistant to high levels of tetracycline either because the tet(l) gene mutated (name of gene might be incorrect but natural tetR resistance gene exists) or the strain we were given was already very resistant. The other possibility is that it was given with a plasmid :: TetR, the last possibility is that our tetracycline is non-functional (photodegradation, others ?). In any case, I doubled and quadrupled concentrations and replated as little biofilm as possible of each of the transformants.
In parallel, I am going to test a simple construct to test efficiency
pVEG-SpoVG-RFP-terminator
bacterial strains were inoculated today.
