Difference between revisions of "Part:BBa K1060003:Experience"

(MeS protein expression experiment)
(System testing)
 
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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.
 
=== System testing ===
 
 
'''Smell Test'''
 
 
https://static.igem.org/mediawiki/2013/8/8e/BBa_K1060003_SmellTest_24h_37C.jpg
 
 
 
To test this device several setups for smell tests where made. In the graph shown here the samples were incubated for 24h at 37°C. Methyl salicylate (MeS) has a wintergreen odor and can be detected by scent. 11 people smelled each sample independently of one another and answered if they could smell MS or not. For more details on the experimental setup and a discussion of the results go [http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/MeS here] under 'smell test'. Chorismate was added to test the salicylate production ([https://parts.igem.org/Part:BBa_J45320 BBa_J45320]) and salicylate to test the wintergreen odor generator ([https://parts.igem.org/Part:BBa_J45120 BBa_J45120]).
 
 
 
'''Smell test 2.0'''
 
 
https://static.igem.org/mediawiki/2013/1/14/Smell_test_KULEUVEN_upgrade.jpg
 
 
Here we tested and compared the activity of the original MIT 2006 Brick ([https://parts.igem.org/Part:BBa_J45700 BBa_J45700] and our Brick under different conditions via a blind smell test (11 participants). The MeS (wintergreen) smell got stronger from 8 to 24 hours incubation time. Growing bacteria at 37 °C also produced a more pronounced smell than those at room temperature. Addition of salicylate positively affected the smell, independent of the incubation time. The chorismate effect became only noticeable after 24 hours.
 
 
We chose to further characterise the MeS generator along with our other Bricks in the pSB1C3 shipping vector to allow comparisons to be made with other Bricks in the registry.
 
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 00:20, 29 October 2013

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.

User Reviews

UNIQc25cea38dd8e121f-partinfo-00000000-QINU UNIQc25cea38dd8e121f-partinfo-00000001-QINU

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Sander Wuyts

The KU Leuven iGEM 2013 team tried to perform a qPCR on this part. It was impossible to remove the original plasmid DNA, after RNA isolation, even after several attempts with different DNase treatments. This problem is probably due to the fact that this part was provided in a high copy number backbone. If you want to perform a qPCR yourself, we recommend you to clone this part in another backbone or in the genome itself.

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