Part:BBa_K2629001:Experience

Experiments were done on a plasmid in which the probe has been inserted, thanks the Gibson technic, in psB1C3-BBa_J04450.

Results of the clonning of the probe into psB1C3-BBa_J04450

This is the alignment between psB1C3-BBa_K2629001 and the probe after probe activation by digestion.

Unfortunately, this is not the result that we expected. In fact, the sequencing shows that the cloning did not work very well for psB1C3-BBa_K2629001. Indeed, 3 mutations can be noted and should not be present.

Test A: Is this part able to detect the target for which it has been designed ?

Unfortunately, results were too heterogenous to bring any conclusions. Indeed, the number of colonies, for the 1:100 and 1:200 ratios, expected was not good enought (there is a possibility that the detector was badly digested and was consequently transformed).

Test B: Is this part able to detect specifically the target for which it has been designed ?

In order to evaluate the specificity of the detector part of BBa_K2629001, different “false target” sequences have been synthesized. An algorithm was made by iGEM Grenoble Alpes 2017 to give random sequences, more or less homologous to the original target with 5%, 15%, 25% and 50% randomly modified pairs of nucleotides (length is kept 36bp).
The algorithm can be found here :

Unfortunately, we did not have the opportunity and the time to carry out these experiments.

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