Team:Paris Saclay/Notebook/August/1

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(→Friday 1st August)
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*Nothing has grown on the control dishes.
*Nothing has grown on the control dishes.
*Failure, nothing has grown on the dishes which contain DY330 with pJBE1
*Failure, nothing has grown on the dishes which contain DY330 with pJBE1
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 +
====New Transformation of electrocompetent cells====
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''by Terry''
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 +
Strain used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)
 +
 +
Make 3 électroporations in cold electroporation cuvettes:
 +
 +
*A control cuvette(without DNA): 50µl of DY330.
 +
*A second cuvette: 50µl of DY330 culture + 2µl of pJBEI-6409 plasmid.
 +
*A third cuvette: 50µl of DY330 culture + 0.5µl of pJBEI-6409 plasmid.
 +
 +
Electroporation : 2500V, 132W, 40µF.
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 +
After that, add 1ml of cold LB in each cuvette and transfer in tubes to incubate during 1 to 2h at 30°C.
 +
 +
Spread on 7 dishes LB + Cm:
 +
 +
*100µl of control (without plasmid)
 +
*50µl of transformed DY330 with 0.5µl of pJBEI-6409
 +
*100µl of transformed DY330 with 0.5µl of pJBEI-6409
 +
*The rest of transformed DY330 with 0.5µl pJBEI-6409
 +
*50µl of transformed DY330 with 2µl of pJBEI-6409
 +
*100µl of transformed DY330 with 2µl of pJBEI-6409
 +
*The rest of transformed DY330 with 2µl pJBEI-6409
 +
 +
Incubate for 2 days at 30°C.

Revision as of 13:28, 5 August 2014

Contents

Friday 1st August

Lab work

A - The frame coli Odor free

Stria on LB dishes

by Romain

Strains used: MG1655 and MG1655Z1, transformed the 30th July, striated on 4 LB dishes per strain.

C - Salicylate Inducible Suppressing System

Liquid Culture

by Fabio

8 liquid cultures with 3ml of LB and 3µl of amp (at 10am - 8°C), all from the colonies made the 31st July. The colonies will be put at 37°C on sunday 3rd in order to have grown colonies on monday.

D - Lemon scent

Results of the Transformation of electrocompetent cells

by Arnaud & Terry

Transformation of electrocompetent cells made the July 29th. Strains used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)

Results:

  • Nothing has grown on the control dishes.
  • Failure, nothing has grown on the dishes which contain DY330 with pJBE1

New Transformation of electrocompetent cells

by Terry

Strain used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)

Make 3 électroporations in cold electroporation cuvettes:

  • A control cuvette(without DNA): 50µl of DY330.
  • A second cuvette: 50µl of DY330 culture + 2µl of pJBEI-6409 plasmid.
  • A third cuvette: 50µl of DY330 culture + 0.5µl of pJBEI-6409 plasmid.

Electroporation : 2500V, 132W, 40µF.

After that, add 1ml of cold LB in each cuvette and transfer in tubes to incubate during 1 to 2h at 30°C.

Spread on 7 dishes LB + Cm:

  • 100µl of control (without plasmid)
  • 50µl of transformed DY330 with 0.5µl of pJBEI-6409
  • 100µl of transformed DY330 with 0.5µl of pJBEI-6409
  • The rest of transformed DY330 with 0.5µl pJBEI-6409
  • 50µl of transformed DY330 with 2µl of pJBEI-6409
  • 100µl of transformed DY330 with 2µl of pJBEI-6409
  • The rest of transformed DY330 with 2µl pJBEI-6409

Incubate for 2 days at 30°C.


Members there:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Arnaud, Fabio, Romain, Sean and Terry.

Back to the calendar

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