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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.
- PCR to amplify Streptavidin without a stop codon needed for the INP-Streptavidin-YFP construct. Gel showed, that nothing was amplified. Repeated the assay with a different PCR programm. Gel showed expected bands.
Inoculation for Western Blot
Overnight inoculation of ISA, ISI, ISD, Streptavidin initial plasmid transformed cells and competent cells.

Sensing-Effector

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.

Nanoparticles

Digestion assays

Culture of MMP2 and gelE from Sensing group