Team:Paris Saclay/Notebook/July/10

From 2014.igem.org

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(2 - Transformation of competent cells E.coli)
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{{Team:Paris_Saclay/notebook_header}}
=Thursday 10th July=
=Thursday 10th July=
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==Lab work==
==Lab work==
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====Plasmid DNA Extraction====
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===1 - Plasmid DNA Extraction:===
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''by Romain and Sean''
''by Romain and Sean''
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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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===The chassis coli Odor free===
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====Transformation of competent cells E.coli====
''by Fabio and Maher''
''by Fabio and Maher''
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BioBricks used:
BioBricks used:
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*J23100 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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*BBa_J23100 (Amp R)(Dilutions: Concentrated, 0 and 10<sup>-1</sup>)
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*J23106 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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*BBa_J23106 (Amp R)(Dilutions: Concentrated, 0 and 10<sup>-1</sup>)
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*J23114 (Amp R)(Dilutions: Concentrated, 0 and 10^-1)
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*BBa_J23114 (Amp R)(Dilutions: Concentrated, 0 and 10<sup>-1</sup>)
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+ pET28a (Kan R)(Dilutions: 0, 10^-1 and 10^-2)
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+ pET28a (Kan R)(Dilutions: 0, 10<sup>-1</sup> and 10<sup>-2</sup>)
(pET28a is a plasmid used to verify that the 4 strains are competent).
(pET28a is a plasmid used to verify that the 4 strains are competent).
Negative control groups:
Negative control groups:
*pET28a: without plasmid, nothing grows.
*pET28a: without plasmid, nothing grows.
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*J23100, J23106, J23114: without the BioBrick, nothing brows.
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*BBa_J23100, BBa_J23106, BBa_J23114: without the BioBrick, nothing grows.
'''Results:'''
'''Results:'''
*pET28a: Nothing grows with the strains DH5α, MG1655, MG1655Z1. But the strain DH5α+ grows well.
*pET28a: Nothing grows with the strains DH5α, MG1655, MG1655Z1. But the strain DH5α+ grows well.
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*J23100, J23106, J23114: The strain DH5α+ grow well and the 3 others strains grow less. They have to be more concentrated.
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*BBa_J23100, BBa_J23106, BBa_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/Protocols/Transformation_of_supercompetent_Ecoli_cells_with_CaCl2 Protocol]
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[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the calendar]
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==Modeling==
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''by Arnaud, Eugene, Juliette and Pierre''
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Discussion around differents modeling possibilities in concertation with Solenne.
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==Photo of the Day==
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[[File:Paris Saclay 10_july.jpg|600px|center]]
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'''Members present:'''
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* Instructors and advisors: Alice, Solenne and Sylvie.
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* Students: Arnaud, Fabio, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain and Sean.
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{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 15:38, 14 October 2014

Contents

Thursday 10th July

Lab work

Plasmid DNA Extraction

by Romain and Sean

Plasmids used:

  • BBa_B0015, dble ter
  • FNR_RBS_amilCP-Ter DH5αZ1

Protocol

The chassis coli Odor free

Transformation of competent cells E.coli

by Fabio and Maher

Competent strains used:

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

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

BioBricks used:

  • BBa_J23100 (Amp R)(Dilutions: Concentrated, 0 and 10-1)
  • BBa_J23106 (Amp R)(Dilutions: Concentrated, 0 and 10-1)
  • BBa_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).

Negative control groups:

  • pET28a: without plasmid, nothing grows.
  • BBa_J23100, BBa_J23106, BBa_J23114: without the BioBrick, nothing grows.

Results:

  • pET28a: Nothing grows with the strains DH5α, MG1655, MG1655Z1. But the strain DH5α+ grows well.
  • BBa_J23100, BBa_J23106, BBa_J23114: The strain DH5α+ grow well and the 3 others strains grow less. They have to be more concentrated.

Protocol

Modeling

by Arnaud, Eugene, Juliette and Pierre

Discussion around differents modeling possibilities in concertation with Solenne.

Photo of the Day

Paris Saclay 10 july.jpg

Members present:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Arnaud, Fabio, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain and Sean.


Back to the calendar