Team:Paris Saclay/Notebook/July/10

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(Difference between revisions)
(→Thursday 10th July)
(→Thursday 10th July)
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol]
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol]
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====2 - Transformation of competent cells E.coli====
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 +
''Fabio & Maher''
 +
 +
Competent strains used:
 +
*DH5α
 +
*MG1655
 +
*MG1655Z1
 +
*DH5α+ (given by Solenne)
 +
(The strains DH5α, MG1655 and MG1655Z1 were prepared by ''Mathias & Mathieu'')
 +
 +
BioBricks used:
 +
*J23100 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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*J23106 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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*J23114 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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+ pET28a (Kan R)(Dilutions: 0, 10^-1 and 10^-2)
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(pET28a is a plasmid used to verify that the 4 strains are competent).
 +
 +
The negative controles:
 +
*pET28a: without plasmid, nothing grows.
 +
*J23100, J23106, J23114: without the BioBrick, nothing brows.
 +
 +
Results:
 +
*pET28a: Nothing grows with the strains DH5α, MG1655, MG1655Z1. But the strain DH5α+ grows well.
 +
*J23100, J23106, J23114: The strain DH5α+ grow well and the 3 others strains grow less. They have to be more concentrated.
 +
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_supercompetent_Ecoli_cells_with_CaCl2 Protocol]
[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]
[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]

Revision as of 07:43, 15 July 2014

Contents

Thursday 10th July

Lab work

1 - Plasmid DNA Extraction:

Romain & Sean

Plasmids used:

  • BBa_B0015, dble ter
  • FNR_RBS_amilCP-Ter DH5αZ1

Protocol

2 - Transformation of competent cells E.coli

Fabio & Maher

Competent strains used:

  • DH5α
  • MG1655
  • MG1655Z1
  • DH5α+ (given by Solenne)

(The strains DH5α, MG1655 and MG1655Z1 were prepared by Mathias & Mathieu)

BioBricks used:

  • J23100 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
  • J23106 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
  • J23114 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)

+ pET28a (Kan R)(Dilutions: 0, 10^-1 and 10^-2) (pET28a is a plasmid used to verify that the 4 strains are competent).

The negative controles:

  • pET28a: without plasmid, nothing grows.
  • J23100, J23106, J23114: without the BioBrick, nothing brows.

Results:

  • pET28a: Nothing grows with the strains DH5α, MG1655, MG1655Z1. But the strain DH5α+ grows well.
  • J23100, J23106, J23114: The strain DH5α+ grow well and the 3 others strains grow less. They have to be more concentrated.

Protocol

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/July/10"