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From 2014.igem.org

Contents 1 Thursday 24th July 1.1 Lab work 1.1.1 B - Construction of the fusion protein (color) 1.1.1.1 Plasmid DNA Extraction 1.1.2 C - Lemon scent 1.1.2.1 Gel electrophoresis of p cola 1.1.2.2 Results of the PCR Targeting 1.1.3 E - The Suppressing Salicylate System 1.1.3.1 Transformation of competent E.coli cells

Thursday 24th July

Lab work

B - Construction of the fusion protein (color)

Plasmid DNA Extraction

by Terry

Biobricks used:

• BBa_K1033927 (pink chromoprotein)
• BBa_K1033910 (yellow chromoprotein)
• BBa_K1033921 (red chromoprotein)
• BBa_K1033922 (red chromoprotein)
• BBa_K1033902 (blue chromoprotein)
• BBa_K731201 (Arabinose inductible araC-pBAD promoter)
• BBa_K762100 (Limonen synthase1 with RBS)
• BBa_K1033925 (pink chromoprotein)
• BBa_K1033905 (purple chromoprotein)
• BBa_K1033913 (orange chromoprotein)
• BBa_K1033929 (blue chromoprotein)
• BBa_J45017 (pchBA synthase (converts chorismate to salicylate))

C - Lemon scent

Gel electrophoresis of p cola

by Sean

p cola used: sample left overnight yesterday resuspended in 30μl of H₂O milliQ.

Protocol

Results

Results of the PCR Targeting

by Romain

PCR targeting made the July 22nd.

Results:

• Nothing has grown on the control dishes.
• Failure, nothing has grown on the clone I dishes.
• Failure, other bacteria have grown on the clone II dishes.

We have to make another PCR.

E - The Suppressing Salicylate System

Transformation of competent E.coli cells

by Fabio

We transformed in DH5α both BioBricks rehydrated yesterday (BBa_J61051 and BBa_K228001).

Samples:

1. BBa_J61051 Concentrated
2. BBa_J61051 Undiluted
3. BBa_K228001 Concentrated
4. BBa_K228001 Undiluted
5. DH5α Control

Protocol

Members present:

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

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