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Part:BBa_K1132031:Experience

Designed by: iGEM Toulouse   Group: iGEM13_INSA_Toulouse   (2013-09-19)
Revision as of 22:48, 4 October 2013 by Nthpham (Talk | contribs) (→‎In vitro characterization of BBa_K1132031)

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In vitro characterization of BBa_K1132031

The basic idea is to perform the switch with a cell lysis containing the recombinase and the plasmid containing the gate to be switched. After incubation, transformation of the plasmid containing the gate allows the quantification of switched versus non switched plasmids. Experiments have been performed as described in the protocol. http://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb

Results

In the absence of recombinase, the colonies are white. As a result, AND gate does not have transcriptional leakage and non-specific recombinases cannot switch the door.

A1.PNG

We tested the AND gate with Bxb1. After incubation with Bxb1, the colonies are still white (this is normal!), therefore visual inspection of the switch was not possible. The only possibility was to sequence independent clones We analyzed five independent colonies by sequencing, all of them were switched.

689px-Switch_AND_1.png

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