Difference between revisions of "Part:BBa K2629001:Experience"

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<h1> Results of the clonning of the probe into psB1C3-BBa_J04450 </h1>
 
<h1> Results of the clonning of the probe into psB1C3-BBa_J04450 </h1>
  
Sequencage
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This is the alignment between psB1C3-BBa_K2629001 and the probe after probe activation by digestion.
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<center>https://static.igem.org/mediawiki/parts/9/93/T--grenoble-alpes--sequencageRESRFP.png </center>
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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.
  
 
<h1> Test A: <I> Is this part able to detect the target for which it has been designed ?</I> </h1><br>
 
<h1> Test A: <I> Is this part able to detect the target for which it has been designed ?</I> </h1><br>

Revision as of 11:05, 9 October 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

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

T--grenoble-alpes--sequencageRESRFP.png

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 ?


T--grenoble-alpes--A1.png

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 :


T--grenoble-alpes--RESalgo.png

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

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