Part:BBa_K2629003: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 BFP gene instead of BBa_J04450 and the probe clonning

This is the alignments realized:

- between psB1C3-BBa_K2629003 and new BFP gene (A).
- between psB1C3-BBa_K2629003 and the probe after probe activation by PCR linearization (B).

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_K2629003, 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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