Revision as of 13:24, 23 September 2018

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

Sequencage

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

mettre schema

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 and tested with the detector. 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 :

These 50%, 75%, 85%, 95% homologous sequences to the target were tested 6 times. Unfortunately, results were too heterogenous to bring any conclusions, notably about the number of false positive CFU (there is a possibility that the detector was badly digested and was consequently transformed) or the supposedly increasing number of colonies. Indeed, transformation includes lot of steps with specific parameters, that all vary a little between each experiment and may explain the lack of reproducibility.

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