Team:Freiburg/Notebook/11 August

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green light receptor

Skypephoned with the iGEM-Team Uppsala

Investigators:Theo, Rüdiger, Jakob

We talked to two guys (Lei Sun and Hamid Gavali) from the igem team uppsala. They gave us some informations and support, quite nice guys.

Informations:

• few Primer sequences
• Touchdown-pcr protocol

Thx a lot for that!!!

blue light receptor

PCR

PCR-Mixture for one Reaction:

For a 50 µl reaction use

What program do you use? Normal PCR with 60°C annealing temperature

To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.

How did you label the PCR-Product, where is it stored and what do you do next?

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME

Precipitator

Tramsformation with inducible Promoter

Investigators: Sophie We didn't get any clones with both Plastic binding domain and Promoter-RBS inside, the Pbd seems to be toxic when expressed.So we want to try if it is possible to get cells with an inducible Promoter and Pbd. In this case we could cultivate the cells and induce expression of the Pbd only shortly before lysis. As our Light-inducible Promoters are not finished we use the IPTG inducible Promoter BBa_J04500 (Plate4, 12A)

PCR

Investigators: Rüdiger

PCR of precipitator.

Primer fw:

• p51
• p52
• p53

Primer dw:

• p54

DNA template: S43

DMSO was used to reduce Tm about 5°C.

For the first a51, a52 and a53 6% DMSO was used. For b51, b52 and b53 9% Dmso was used. For c51, c52 and c53 12% Dmso was used.

Picking colonies

Investigators: Rüdiger

5 colonies were picked and incubated overnight in LB amp medium.