Difference between revisions of "Part:BBa K2629000:Experience"
Line 11: Line 11:
  
 
mettre schema
 
mettre schema
 +
 +
<p> 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). </p>
  
 
<h1> Test B: <I> Is this part able to detect specifically the target for which it has been designed ? </I> </h1>
 
<h1> Test B: <I> Is this part able to detect specifically the target for which it has been designed ? </I> </h1>
  
  
−
<p> 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.  
+
<p> 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).  <br>
 
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).  <br>
 
The algorithm can be found here : </p><br>
 
The algorithm can be found here : </p><br>
Line 21: Line 23:
 
<center>https://static.igem.org/mediawiki/parts/4/4b/T--grenoble-alpes--algo.png</center>
 
<center>https://static.igem.org/mediawiki/parts/4/4b/T--grenoble-alpes--algo.png</center>
  
−
<p>These 50%, 75%, 85%, 95% homologous sequences to the target were tested 6 times.
+
<p> Unfortunately, we did not have the opportunity and the time to carry out these experiments. </p>
−
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. </p>
+
  
 
===User Reviews===
 
===User Reviews===

Revision as of 14:03, 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

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_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 :


T--grenoble-alpes--algo.png

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

User Reviews

UNIQ6358eb0b39a66c05-partinfo-00000000-QINU UNIQ6358eb0b39a66c05-partinfo-00000001-QINU