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From 2014.igem.org

Contents 1 Thursday 10th July 1.1 Lab work 1.1.1 1 - Plasmid DNA Extraction: 1.1.2 2 - Transformation of competent cells E.coli

Thursday 10th July

Lab work

1 - Plasmid DNA Extraction:

by Romain and Sean

Plasmids used:

• BBa_B0015, dble ter
• FNR_RBS_amilCP-Ter DH5αZ1

Protocol

2 - 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:

• 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

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