Team:TUDelft/23 July 2009

Tim Weenink

Details of the following to be provided tomorrow:

grew up *S in LC

looked at gel

Did !A assembly again with double the amount of backbone (and rest from fridge). Did it with both DH5alpha and homemade top10 chemically competent cells, to compare transformation efficiencies. Also the right concentration of K was used

PCR'd 8 colonies from my transformation plates with VF2 and I-SceI Reverse primers to see if there was incorporation of the I-SceI restriction site

isolated plasmid from *S

Sriram

RThe gel was run for another 1 hour and 10 minutes and the TBE buffer was stained with 20&mico;l of Green gel stain and the bands were viewed to check whether we have the right biobricks. By this we confirmed that all the amplified biobricks to be used in Delay were extracted well and they are pure. [The gel image will be uploaded tomorrow.]

For preparing glycerol stocks and plasmid DNA extraction the 7 biobrick colonies [R0010, R0040, J23008, J23031, E1010, B0034, B0015, K081013] from the LB agar plates and 2 composite biobricks received from iGEM HQ were cultured in 5 ml tubes with 1xAmp. The 7 biobricks were recultured since we anticipate we may need more DNA for assembly for parts like RBS, Double terminator, etc. and also glycerol stocks were not made for these biobricks.

The 2 composite biobricks were also streaked in LB Agar plates with 1xAmp for future use.