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Team:Berkeley/Project/Construction
From 2012.igem.org
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Revision as of 05:42, 2 October 2012
We chose to use Golden Gate Assembly because it allowed us to either create single-pot libraries or the interchange of combinatorially expand the number of multi-gene devices with smaller constituent parts. Golden Gate Assembly (GGA) is powered by Type IIs restriction enzymes, which cut distal to their recognition sites. Because of this, we have full control over the resulting 4bp overhangs, giving us a theoretical choice between 4^4, or 256 different overhangs. In practice, we reduced this set, eliminating cases where three out of the four matched to minimize chances of mis-annealing.
An example type IIs restriction enzyme, BsmBI.
The basic, fundamental unit of our GGA is the part plasmid, shown below. We
