Difference between revisions of "Part:BBa K4174002:Experience"
(→‎William and Mary iGEM 2022)
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====William and Mary iGEM 2022====
 
====William and Mary iGEM 2022====
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To test the effectiveness of our sfGFP construct (BBa_K4174002), our team transformed the original MIT iGEM 2006 construct, our improved sfGFP construct, and our improved RFP construct (BBa_K4174001) in <i>E. coli</i> NEB5α in a plate reader. The various transformants were grown at 37°C. For green fluorescence, we used an excitation value of 485 and an emission value of 528.  For red fluorescence, we used an excitation value of 584 and an emission value of 610. The values for green fluorescence are reported below. For information about red fluorescence, see parts page BBa_K4174001.
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To test the effectiveness of our osmY-sfGFP construct (BBa_K4174002), our team transformed the original MIT iGEM 2006 osmY construct (BBa_J45995), our improved osmY-sfGFP construct, and our improved osmY-mRFP1 construct (BBa_K4174001) in <i>E. coli</i> NEB5-α in a plate reader. The various transformants were grown at 37°C with continuous shaking. For green fluorescence, we used an excitation wavelength of 485 nm and an emission wavelength of 528 nm. The values for green fluorescence are reported below. For information about red fluorescence, see parts page BBa_K4174001.
  
 
https://static.igem.wiki/teams/4174/wiki/normalized-green-fluorescence-graph-final-larger.png
 
https://static.igem.wiki/teams/4174/wiki/normalized-green-fluorescence-graph-final-larger.png
  
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As seen in the graph above, both the sfGFP and MIT GFP constructs enter stationary phase right before 14 hours, but our improved sfGFP circuit is much more fluorescent. The other constructs are our RFP construct and untransformed <i>E. coli</i> cells, both of which serve as negative controls for green fluorescence.
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As seen in the graph above, both the osmY-sfGFP (BBa_K4174002) and MIT iGEM 2006 osmY (BBa_J45995) constructs enter stationary phase right before 14 hours, but our improved osmY-sfGFP construct (BBa_K4174002) is much more fluorescent. The other constructs are our osmY-mRFP1 construct (BBa_K4174001) and untransformed NEB5-α <i>E. coli</i> cells, both of which serve as negative controls for green fluorescence.
  
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https://static.igem.wiki/teams/4174/wiki/improveapart-smaller.png
 
https://static.igem.wiki/teams/4174/wiki/improveapart-smaller.png
  
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As seen in the image above, qualitative results reveal that our improved constructs are more fluorescent than the original construct. Here, sfGFP is on the far right, and is visibly more brightly green that the original GFP construct.
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As seen in the image above, qualitative results reveal that our improved constructs are more fluorescent than the original construct (BBa_J45995). Here, a culture of bacterial cells engineered with our osmY-sfGFP construct (BBa_K4174002) is on the far right, and is visibly more green that the culture engineered with the original MIT 2006 iGEM osmY construct (BBa_J45995).
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===User Reviews===
 
===User Reviews===

Revision as of 19:13, 10 October 2022

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_K4174002

William and Mary iGEM 2022

To test the effectiveness of our osmY-sfGFP construct (BBa_K4174002), our team transformed the original MIT iGEM 2006 osmY construct (BBa_J45995), our improved osmY-sfGFP construct, and our improved osmY-mRFP1 construct (BBa_K4174001) in E. coli NEB5-α in a plate reader. The various transformants were grown at 37°C with continuous shaking. For green fluorescence, we used an excitation wavelength of 485 nm and an emission wavelength of 528 nm. The values for green fluorescence are reported below. For information about red fluorescence, see parts page BBa_K4174001.

normalized-green-fluorescence-graph-final-larger.png

As seen in the graph above, both the osmY-sfGFP (BBa_K4174002) and MIT iGEM 2006 osmY (BBa_J45995) constructs enter stationary phase right before 14 hours, but our improved osmY-sfGFP construct (BBa_K4174002) is much more fluorescent. The other constructs are our osmY-mRFP1 construct (BBa_K4174001) and untransformed NEB5-α E. coli cells, both of which serve as negative controls for green fluorescence.


improveapart-smaller.png


As seen in the image above, qualitative results reveal that our improved constructs are more fluorescent than the original construct (BBa_J45995). Here, a culture of bacterial cells engineered with our osmY-sfGFP construct (BBa_K4174002) is on the far right, and is visibly more green that the culture engineered with the original MIT 2006 iGEM osmY construct (BBa_J45995).

User Reviews

UNIQ0db063c1de030c57-partinfo-00000000-QINU UNIQ0db063c1de030c57-partinfo-00000001-QINU