Part:BBa_K2629000: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

In parallel of the experiments, we sent our newly designed probe to sequencing. Unfortunately, the insert was not found in the sequence when alignment was done, the size was not the right one and the only thing that we could find was the sequence of BBa_J04450.

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 badly 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_K2629000, 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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