Team:Aberdeen Scotland/WetLab/andgate/cloning

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University of Aberdeen iGEM 2009

Cloning Strategy

As our individual modules would be ideally combined at a later point, we had to make sure that our final vector would be compatible with both the quorum sensing module and LacI-Lach module, thus we settled on pSB3T5 as our final vector.

University Aberdeen 2009 ANDgateStep1.PNG

Our first step was to join CI repressor with a ribosome binding site, green fluorescence protein and the double terminator. For this step we decided to use biobricks C0051, E0840 and clone into the pSB1AC3. As CI repressor gene was upstream we digested it with EcoRI and SpeI restriction enzymes. The downstream part was digested with XbaI and PstI, therefore vector was cut with EcoRI and PstI. Our experience with plasmid digests led us to use high copy number. This initial step was the creation of biobrick <a href="http://partsregistry.org/Part:BBa_K182100">K182100</a>.

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