Difference between revisions of "Part:BBa K2629003:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
how you used this part and how it worked out. | how you used this part and how it worked out. | ||
− | ===Applications of | + | ===Applications of BBa_K2629001=== |
+ | |||
+ | |||
+ | <p>Experiments were done on a plasmid in which the probe has been inserted, thanks the Gibson technic, in psB1C3-BBa_J04450. </p> | ||
+ | |||
+ | <h1> Test A: <I> Is this part able to detect the target for which it has been designed ?</I> </h1><br> | ||
+ | |||
+ | mettre schema | ||
+ | |||
+ | <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. | ||
+ | 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> | ||
+ | |||
+ | <center>https://static.igem.org/mediawiki/parts/9/96/T--grenoble-alpes--RESalgo.png</center> | ||
+ | |||
+ | <p>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. </p> | ||
===User Reviews=== | ===User Reviews=== | ||
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<I>Username</I> | <I>Username</I> | ||
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Revision as of 10:38, 12 September 2018
_NOTOC__
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Contents
Applications of BBa_K2629001
Experiments were done on a plasmid in which the probe has been inserted, thanks the Gibson technic, in psB1C3-BBa_J04450.
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.
User Reviews
UNIQab7c52d27b20d983-partinfo-00000000-QINU UNIQab7c52d27b20d983-partinfo-00000001-QINU