Team:EPF Lausanne/Calendar/15 September 2013


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Taxi.Coli: Smart Drug Delivery iGEM EPFL


Cell Surface Display

New Strategy
- Optimization of PCR using the new primers designed to amplify the different component that will be needed to clone the following constructs : INP_YFP_Streptavidin and INP_streptavidin_YFP.
- Beginning of gibson reactions involving the previously amplified DNA sequences.
Inoculation for Western Blot
Overnight inoculation of ISA, ISI, ISD, Streptavidin initial plasmid transformed cells and competent cells.


Analysis of the sequencing results of the three promoters (sensing-module)
-The sequencing results were rather promising except for the Hya promoter. The sample that I send did not contain it. So I purified one in order to send it again on Tuesday. This sample was from one of the assays. I send this sample because it did not turn green when I added arabinose indicating that the Arabinose inducible promoter had effectively been removed and hopefully replaced by the hya promoter.

Functional assays for the three promoters (sensing-module)
-I again tried to grow the bacteria in an LB medium to which I added different amounts of either 10X MOPS or 10X HEPES buffer. But again the cultures did not turn green.

MinPrep of the hya-plasmid
-Because the sequencing results showed that the plasmid I sent did not contain the hya promoter I picked another colony that I incubated and from which I then isolated the plasmid again by a MiniPrep.


Digestion assays

Culture of MMP2 and gelE from Sensing group