Team:Paris Bettencourt/Notebook/Trojan Horse/Tuesday 2nd July.html

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     <em>Aude and Vincent</em>
     <em>Aude and Vincent</em>
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     - plating 3 different quantity of cells (20ul, 50uL, 100uL) respectively on Cm and Kan plates (dilution 1000x for the antibiotics)
     - plating 3 different quantity of cells (20ul, 50uL, 100uL) respectively on Cm and Kan plates (dilution 1000x for the antibiotics)
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    03/07
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    <em>Aude and Vincent</em>
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    Transfo results :
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            negative controls are negative.
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            Transformed cells grew on plates
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            in the evening launch of 5mL Lb cultures from clones on the plates
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    <strong>Igem Buffer for chemical competent cells</strong>
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    Igem protocol for chemical competent cells
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    <strong>Launch overnight culture of NEB turbo</strong>
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    5mL LB inoculated with a clone from plate of drug team
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Latest revision as of 12:56, 29 September 2013

Trojan Horse

2nd July


Aude and Vincent

glycerol stock

from overnight culture of MG1655-6300 O/N : T001

  • Centrifuge 4000rpm, 10 minutes,

  • take out liquid

  • resuspend cells in 1mL glycerol, 2mL LB

  • separate in two cryotubes, one for the -80°C, one for the -20°C

Electroporation

- making MG1655-6300 competent (Aude’s protocol for electrocompetent cells & electroporation [ref needed])

- test of the competent cells (negative

-transforming with pCOLADuet-1 : Ec2 =>

-transforming with pACYCDuet-1 : Ec2 =>

- plating 3 different quantity of cells (20ul, 50uL, 100uL) respectively on Cm and Kan plates (dilution 1000x for the antibiotics)

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