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TechnologyBioinformatics 
CLONING
Expression
Purification
Crystallisation
Structure
Our current cloning strategy relies on the Gateway system of ligation independent cloning. We use a pDEST14 destination vector, and add a N-terminal His tag and TEV cleavage site to the protein of interest. We are also experimenting with a non-cleavable C terminal His tag (see below). 
We are also developing automated techniques for cloning, based on a Hamilton Microlab Star. This robot will carry out the following steps of the cloning process: 1) Automated PCR 2) E-gel analysis for DNA and protein 3) DNA cleanup 4) BP and LR recombination 5) Heat-shock mediated transformation 6) Colony picking 7) Cell inoculation 8) Incubation 9)DNA mini-prep With this technology we will be able to clone and transform 96 constructs a week. 
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