Difference between revisions of "Part:BBa K546001:Experience"

(→‎User Reviews)
(→‎User Reviews)
Line 14: Line 14:
 
<I>Youri van Nuland</I>
 
<I>Youri van Nuland</I>
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
 +
 +
==Protocol==
 +
  
 
Functionality of this part was tested by treating AHL mixtures with strains expressing this part. Liquid cultures were grown overnight, 1 mL aliquots of these cultures were spun down and resuspended in varying concentrations of AHL in PBS (to start the AHL degradation reaction). This was also done with a culture that contains BBa_K546002 (where aiiA is replaced with GFP).  
 
Functionality of this part was tested by treating AHL mixtures with strains expressing this part. Liquid cultures were grown overnight, 1 mL aliquots of these cultures were spun down and resuspended in varying concentrations of AHL in PBS (to start the AHL degradation reaction). This was also done with a culture that contains BBa_K546002 (where aiiA is replaced with GFP).  
  
−
After a 3h incubation, the supernatant was collected and subjected to a reporter strain: BBa_K546002. The result is shown below.  
+
After a 3h incubation, the supernatant was collected and subjected to a reporter strain: BBa_K546002. 200 µL samples of reporter strain/AHL mixtures were analyzed with a Molecular Devices Spectramax M2 spectrophotometer, detecting GFP. The result is shown below.  
 +
 
  
 
[[Image: AHL_degradation_by_AiiA_reported_by_BBa_K546002correct.png]]
 
[[Image: AHL_degradation_by_AiiA_reported_by_BBa_K546002correct.png]]
  
 
This part is functional, as AHL concentrations drop after incubation with <I>E. coli</I> transformed with this part. The GFP expression is much higher in the non treated mixtures, namely.
 
This part is functional, as AHL concentrations drop after incubation with <I>E. coli</I> transformed with this part. The GFP expression is much higher in the non treated mixtures, namely.
 +
 +
<I>Team Wageningen_UR 2011</I>
 
|};
 
|};
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_K546001 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K546001 EndReviews</partinfo>

Revision as of 09:56, 21 September 2011

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.

Applications of BBa_K546001

User Reviews

UNIQ9524ce1715f3d7ee-partinfo-00000000-QINU

No review score entered. Youri van Nuland

Protocol

Functionality of this part was tested by treating AHL mixtures with strains expressing this part. Liquid cultures were grown overnight, 1 mL aliquots of these cultures were spun down and resuspended in varying concentrations of AHL in PBS (to start the AHL degradation reaction). This was also done with a culture that contains BBa_K546002 (where aiiA is replaced with GFP).

After a 3h incubation, the supernatant was collected and subjected to a reporter strain: BBa_K546002. 200 µL samples of reporter strain/AHL mixtures were analyzed with a Molecular Devices Spectramax M2 spectrophotometer, detecting GFP. The result is shown below.


AHL degradation by AiiA reported by BBa K546002correct.png

This part is functional, as AHL concentrations drop after incubation with E. coli transformed with this part. The GFP expression is much higher in the non treated mixtures, namely.

Team Wageningen_UR 2011

;

UNIQ9524ce1715f3d7ee-partinfo-00000002-QINU