Difference between revisions of "Part:BBa K2629003:Experience"

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<h1> Results of the clonning of the BFP gene instead of BBa_J04450 and the probe clonning </h1>
 
<h1> Results of the clonning of the BFP gene instead of BBa_J04450 and the probe clonning </h1>
  
sequencage
+
This is the alignments realized: <br>
  
 +
- between psB1C3-BBa_K2629003 and new BFP gene (A). <br>
 +
- between psB1C3-BBa_K2629003 and the probe after probe activation by PCR linearization (B).  <br>
 +
 +
 +
<center>https://static.igem.org/mediawiki/parts/d/dc/T--grenoble-alpes--sequencageRESBFP.png</center>
  
 
<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 11:25, 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 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).


T--grenoble-alpes--sequencageRESBFP.png

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_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 :


T--grenoble-alpes--RESalgo.png

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

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