Team:Paris Liliane Bettencourt/Notebook/2011/08/02/

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(Difference between revisions)
(→Adrien Lhomme-Duchadeuil)
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|-
|-
|tube 1: poration control no DNA (12,5 kV: ARC)
|tube 1: poration control no DNA (12,5 kV: ARC)
-
*CTL <pow>+</pow>: carpet
+
*CTL <sup>+</sup>: carpet
-
*CTL <sub>-</sub> with Cm (5 and 30 µg/mL): none
+
*CTL <sup>-</sup> with Cm (5 and 30 µg/mL): none
-
*CTL <sub>-</sub> with TetR (5 and 12,5 µg/mL): carpet
+
*CTL <sup>-</sup> with TetR (5 and 12,5 µg/mL): carpet
|tube 2: poration control no DNA (10kV: OK)
|tube 2: poration control no DNA (10kV: OK)
-
*CTL <sub>+</sub>: carpet  
+
*CTL <sup>+</sup>: carpet  
-
*CTL <sub>-</sub> with Cm: none
+
*CTL <sup>-</sup> with Cm: none
-
*CTL <sub>-</sub> with TetR: none
+
*CTL <sup>-</sup> with TetR: none
|tube 3: pHM3::TetR (12,5 kV: OK)
|tube 3: pHM3::TetR (12,5 kV: OK)
* TetR(5 and 12,5 µg/mL): carpet
* TetR(5 and 12,5 µg/mL): carpet
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|tube 10:  S12::Cm (10 kV: OK)
|tube 10:  S12::Cm (10 kV: OK)
* Cm 5 µg/mL: 1 colonie
* Cm 5 µg/mL: 1 colonie
-
| CTL <sub>--</sub>: no bacteria
+
| CTL <sup>--</sup>: no bacteria
* no growth
* no growth
|}
|}

Revision as of 10:36, 2 August 2011

Adrien Lhomme-Duchadeuil

First, an estimation of how much antibiotics is necessary for Bacillus subtilis. Appropriate concentration in antibiotics

  • Chloramphemicol : 5-10 µg/mL
  • Spec : 50-100 µg/mL
  • Tetracyclin: 25 µg/mL
  • Kanamycin: 5µg/mL
  • Evm(?): 1µg/mL + 25 µg/mL lincomycin (Verify spelling)

Results of electroporation: IT WORKED after three trials!

  • Positive control worked
  • Negative controls didn't show any colonies except when put in the presence of TetR (qty:12,5 µg/mL or 5µg/mL) which was insufficient => I will re-plate the improbable survivors on plates of different tetracyclin concentrations to test resistance.
  • Cm resistant strain have survived in low quantities at both 30 and 5 µg/mL of antibiotics
  • It seems that even cuvettes that had an electric arc were able to produce a couple colonies, which means that it doesn't anihilate all the bacteria.
  • To come: protocol and table of results
Table of resutls of electroporation using Cao et al. method
tube 1: poration control no DNA (12,5 kV: ARC)
  • CTL +: carpet
  • CTL - with Cm (5 and 30 µg/mL): none
  • CTL - with TetR (5 and 12,5 µg/mL): carpet
tube 2: poration control no DNA (10kV: OK)
  • CTL +: carpet
  • CTL - with Cm: none
  • CTL - with TetR: none
tube 3: pHM3::TetR (12,5 kV: OK)
  • TetR(5 and 12,5 µg/mL): carpet
tube 4: pHM3::TetR (10 kV: OK)
  • TetR (12,5 µg/mL): carpet
tube5: pHM3::TetR (12,5 kV: ARC)
  • TetR (5 and 12,5 µg/mL): carpet
tube 6: pHM3::TetR (10 kV: OK)
  • TetR (5 µg/mL): carpet
tube 7: S12::Cm (12,5 kV: ARC)
  • Cm 5 µg/mL: 1 colony
  • Cm 30 µg/mL: 1 colony
tube 8: S12::Cm (10 kV: OK)
  • Cm 30 µg/mL: 4 colonies
tube 9: S12::Cm (12,5 kV: OK)
  • Cm 30 µg/mL: 9 colonies
  • Cm 5 µg/mL: 4 colonies
tube 10: S12::Cm (10 kV: OK)
  • Cm 5 µg/mL: 1 colonie
CTL --: no bacteria
  • no growth
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