EPF-Lausanne/17 July 2008

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Results of the transformation of yesterday : All cells grew very well, except the Top10 A (X) and the Top10 - (X).

We did glycerol stocks for the pUC19 and pHD1 and also minipreps of these plasmids. Concentrations were 111.1 and 110.0 for the pUC19 and 126.8 and 70.1 for the pHD1.

We began a new transformation, following exactly the iGEM protocol. We took our Top10 cells, sample B. The iGEM parts used are the 4H 1018 and 8H1003. There are parts that will not be used in our project, then we used them just to practise and see the transformation efficiency. We did two transformations for each part : One part diluted in 5microliter of TE (2microliter of it was used) and the other one diluted in 15microliter of TE(6microliter of it was used). A control was made with the pUC19.

We did a digestion of our transformed cells to verify good incorporation of DNA. We used Ste1 for PDH1. This gave us 3 cuts -> 3 strands. For pLD we used Ste1 and EcoR1 to get a total of 3 cuts again. After gel electrophoresis we obtained the desired bands.