Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-16-2014

From 2014.igem.org

(Difference between revisions)
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<li> Ampicilin
<li> Ampicilin
<li> Tetracyclin
<li> Tetracyclin
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</ul> <br>
+
</ul>  
(The last one was the control of the growth of our bacteria without antibiotics)
(The last one was the control of the growth of our bacteria without antibiotics)
We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C.<br>
We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C.<br>
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<li>Top10
<li>Top10
<li>DH5a
<li>DH5a
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</ul><br>
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</ul>
<u>From plates:</u>
<u>From plates:</u>
<ul>
<ul>
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<li>Top10 transformed with pQexp
<li>Top10 transformed with pQexp
<li>DH5a pyr tranformed with pMK2
<li>DH5a pyr tranformed with pMK2
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</ul><br>
+
</ul>
Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection.
Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection.
For each medium we make a negative contrôle without bacteria.
For each medium we make a negative contrôle without bacteria.

Revision as of 17:38, 25 August 2014

Picture

Tests of antibiotics' stocks
Six plates of LB agar were made. Five of them contained one of those antibiotics in the dilution 1:1000:

  • Chloramphénicol
  • Kanamycin
  • Erythromycin
  • Ampicilin
  • Tetracyclin
(The last one was the control of the growth of our bacteria without antibiotics) We sowed 10µL of a liquid LB culture of Bl21, and we let them incubate overnight at 37°C.
Survivability tests
Two plates of MB 1X and M9 1X were made and divised in two parts. Each plate was sowed with 5µL of a liquid MB 1X culture of Pseudovibrio denitrificans(dated 12th August) in a part and with 5µL of a liquid M9 1X culture Pseudovibrio denitrificans (dated 12th August) in the other part.
Pre-cultures
Cultures in MB 1X and M9 1X of Pseudovibrio were passed by a 1/10 dilution and let incubate overnight at 30°C. New cultures of different E.Coli were also started from glycerol or plate and let incubated overnight at 37°C.
From glycerol stocks:
  • Bl21
  • Top10
  • DH5a
From plates:
  • DH5a tranformed with pCB1C3
  • Top10 transformed with pQexp
  • DH5a pyr tranformed with pMK2
Bacteria tranformed with plasmid were cultivated with the corresponding antibiotic for the selection. For each medium we make a negative contrôle without bacteria.

Aug 16

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