Team:Newcastle/ Stochastic Switch cloning strategy

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We also need to clone the brick into a BioBrick compatible vector for submission to MIT. Digesting the fragment with EcoR1 and Pst1 and ligation into a EcoR1 and Pst1 cut biobrick vector will do this.
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===Cloning strategies: Degradation controller===
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Revision as of 16:41, 21 October 2009

Stochastic Switch Cloning Strategy

We have the following cloning strategies for testing our construct.

Cloning strategies: Stochastic switch

The following is the clone manager file for our synthesised brick; using clone manager it is easy to simulate digests and ligation reactions, in order to make sure the enzymes you use in the will give the result you expect.

Team NewcastleClone manager Synthesised.png


The stochastic brick fragment is to be cloned into the integration vector pGFP-rrnB. This is because plasmid DNA is very unstable in bacillus. From pGFP-rrnb it will integrate into the 168 chromosome at amyE. We can clone the fragment by cutting pGFP-rrnB with EcoRI and NheI, cutting the fragment with EcoRI and NheI, and ligating.

pGFP-rrnb

We also need to clone the brick into a BioBrick compatible vector for submission to MIT. Digesting the fragment with EcoR1 and Pst1 and ligation into a EcoR1 and Pst1 cut biobrick vector will do this.

Cloning strategies: Degradation controller



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