Difference between revisions of "Part:BBa J45995:Experience"
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====William and Mary iGEM 2022==== | ====William and Mary iGEM 2022==== | ||
− | To test the effectiveness of our improved parts, our team grew the original MIT 2006 construct, our improved sfGFP construct, and our improved | + | |
+ | <b>BBa_K4174001</b> | ||
+ | |||
+ | To test the effectiveness of our improved parts, our team grew the original construct (BBa_J45995), our improved <i>mRFP1</i> construct, and our improved <i>sfGFP</i> construct (BBa_K4174001) in <i>E. coli</i> NEB5α in a plate reader. They were grown at 37°C using continuous shaking. For red fluorescence measurements, we used an excitation value of 584 nm and an emission value of 610 nm. For green fluorescence measurements, we used an excitation value of 485 nm and an emission value of 528 nm. The values for red fluorescence are reported below. For information about green fluorescence measurements, see parts page BBa_K4174002. | ||
+ | |||
+ | https://static.igem.wiki/teams/4174/wiki/improve-a-part-red-fluorescence-graph.png | ||
+ | |||
+ | Based on the graph above, the bacterial cells engineered with our <i>mRFP1</i> construct appear to have entered stationary phase around 16 hours. As seen in the graph, our <i>mRFP1</i> construct produces more red fluorescence than the original circuit. The other measurements taken are for our <i>sfGFP</i> construct and untransformed <i>E. coli</i> NEB5α cells, both of which serve as negative controls for red fluorescence. | ||
+ | |||
+ | https://static.igem.wiki/teams/4174/wiki/improveapart-smaller.png | ||
+ | |||
+ | As seen in the image above, qualitative results reveal that our <i>mRFP1</i> construct produces more red fluorescence than the original construct. Here, our mRFP1 construct is on the far left, and is visibly more red that the original GFP construct. | ||
+ | |||
+ | <b>BBa_K4174002</b> | ||
+ | To test the effectiveness of our improved parts, our team grew the original MIT 2006 construct, our improved sfGFP construct (BBa_K4174002), and our improved mRFP1 construct (BBa_K4174001) in <i>E. coli</i> NEB5α in a plate reader. They were grown at 37°C using continuous shaking. 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. | ||
https://static.igem.wiki/teams/4174/wiki/normalized-green-fluorescence.png | https://static.igem.wiki/teams/4174/wiki/normalized-green-fluorescence.png |
Revision as of 21:42, 8 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_J45995
Stationary phase dependent fluorescence.
User Reviews
UNIQ63230eaf9223f456-partinfo-00000000-QINU
••••• |
BBa_J45995 produced fluorescence only in stationary phase. |
UNIQ63230eaf9223f456-partinfo-00000003-QINU
Characterization
Transcriptional control of GFP generator
[Note: BBa_J45995 is a composite part of BBa_J45992 and BBa_E0840.]
William and Mary iGEM 2022
BBa_K4174001
To test the effectiveness of our improved parts, our team grew the original construct (BBa_J45995), our improved mRFP1 construct, and our improved sfGFP construct (BBa_K4174001) in E. coli NEB5α in a plate reader. They were grown at 37°C using continuous shaking. For red fluorescence measurements, we used an excitation value of 584 nm and an emission value of 610 nm. For green fluorescence measurements, we used an excitation value of 485 nm and an emission value of 528 nm. The values for red fluorescence are reported below. For information about green fluorescence measurements, see parts page BBa_K4174002.
Based on the graph above, the bacterial cells engineered with our mRFP1 construct appear to have entered stationary phase around 16 hours. As seen in the graph, our mRFP1 construct produces more red fluorescence than the original circuit. The other measurements taken are for our sfGFP construct and untransformed E. coli NEB5α cells, both of which serve as negative controls for red fluorescence.
As seen in the image above, qualitative results reveal that our mRFP1 construct produces more red fluorescence than the original construct. Here, our mRFP1 construct is on the far left, and is visibly more red that the original GFP construct.
BBa_K4174002 To test the effectiveness of our improved parts, our team grew the original MIT 2006 construct, our improved sfGFP construct (BBa_K4174002), and our improved mRFP1 construct (BBa_K4174001) in E. coli NEB5α in a plate reader. They were grown at 37°C using continuous shaking. 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.
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 E. coli cells, both of which serve as negative controls for green fluorescence.
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.