Difference between revisions of "Team:Pasteur Paris/Results"
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| + | <p><em>Gibson assembly: </em></p> | ||
| + | <p>- BioBricks submitted to be BioBrick registry: </p> | ||
| + | <p>- Bba_K1622000: assembly of the Glycoaldehyde Dehydrogenase (BBa_K936011) and the Glycoaldehyde Reductase (BBa_K936023) in the plasmid PSB1C3 :</p> | ||
| + | <p> - Bba_K1622001: assembly of the NB-Esterase (BBa_K808030) and the TPA transporteur (BBa_K808007) </p> | ||
| + | <p> </p> | ||
| + | <p><em>Interlab Study: </em></p> | ||
| + | <p>- Transformation in DH5-alpha of each promoter (BBa_J23101, BBa_J23106, BBa_J23117) and the controls GFP (BBa_I13405) and WT. </p> | ||
| + | <p>- qPCR of the transformed cells to determinate the plasmid copy number per strain. </p> | ||
| + | <p>- fluorescence test of the GFP expression and determination of the fluorescence per plasmid.</p> | ||
| + | <p> </p> | ||
| + | <p> <em>pNP assay: </em></p> | ||
| + | <p>- Insertion of the NB-Esterase (BBa_K808030) in the plasmid pDG011</p> | ||
| + | <p> - Transformation of BAP1 with the construct NB-Esterase (Bba_K808030) in the plasmid pDG011 </p> | ||
| + | <p>- Isolation of three clones who have our construct NB-Esterase (Bba_K808030) in the plasmid pDG011. </p> | ||
| + | <p>- Demonstration of the degradation of the 4-NitroPhenyl Butyrate by our modified BAP1 with NB-Esterase in the pDG011 plasmid. </p> | ||
| + | <p> </p> | ||
| + | <p><em>TPA :</em></p> | ||
| + | <p>- Test of the TPA toxicity thanks to a variation of the TPA concentration.</p> | ||
| + | <p> - Solubilization of TPA and amelioration of the method. </p> | ||
| + | <p> </p> | ||
| + | <p> </p> | ||
| + | <p><strong>Problems: </strong></p> | ||
| + | <p> Gibson assembly: the exonuclease contained in the Master Mix uncovered the Lox Recombination sites because they were too close to the overlapping ends. </p> | ||
| + | <p><em>Interlab Study: </em></p> | ||
| + | <p>Different strain for qPCR and Fluorescence test/ Modeling:</p> | ||
| + | <p> Because of the absence of experimental results, we can't model our enzymatic system. </p> | ||
| + | <p><em>CRE:</em> - the yeast assembly did not work. pNP assay: - Two of our tests are less decisive because we didn't wait for a long time to see a real difference enter control and modified bacteria. - transformation of the construct very hard.</p> | ||
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Revision as of 03:57, 19 September 2015
Results
pNP-Assay
Estimation of the activity of NB-Esterase, slowest enzyme of our degradation chain
TPA toxicity
- Assessment of the toxicity
- Determination that TPA is not degraded
Operon assembly
- Succesful assembly of 2 of our 4 gene clusters (BBa_K1622000 and BBa_K1622001).
- Optimization of DNA sequences for E.coli.
Interlab Study
- Successful building of the 3 devices
- Characterization of the 3 devices using a Tecan micro-plate reader.
- Quantification of the number of Plasmids in each bacteria.
- Determination of the best Promoter
Here you can describe the results of your project and your future plans.
What should this page contain?
- Clearly and objectively describe the results of your work.
- Future plans for the project
- Considerations for replicating the experiments
Project Achievements
You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.
- A list of linked bullet points of the successful results during your project
- A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.
Inspiration
See how other teams presented their results.
Gibson assembly:
- BioBricks submitted to be BioBrick registry:
- Bba_K1622000: assembly of the Glycoaldehyde Dehydrogenase (BBa_K936011) and the Glycoaldehyde Reductase (BBa_K936023) in the plasmid PSB1C3 :
- Bba_K1622001: assembly of the NB-Esterase (BBa_K808030) and the TPA transporteur (BBa_K808007)
Interlab Study:
- Transformation in DH5-alpha of each promoter (BBa_J23101, BBa_J23106, BBa_J23117) and the controls GFP (BBa_I13405) and WT.
- qPCR of the transformed cells to determinate the plasmid copy number per strain.
- fluorescence test of the GFP expression and determination of the fluorescence per plasmid.
pNP assay:
- Insertion of the NB-Esterase (BBa_K808030) in the plasmid pDG011
- Transformation of BAP1 with the construct NB-Esterase (Bba_K808030) in the plasmid pDG011
- Isolation of three clones who have our construct NB-Esterase (Bba_K808030) in the plasmid pDG011.
- Demonstration of the degradation of the 4-NitroPhenyl Butyrate by our modified BAP1 with NB-Esterase in the pDG011 plasmid.
TPA :
- Test of the TPA toxicity thanks to a variation of the TPA concentration.
- Solubilization of TPA and amelioration of the method.
Problems:
Gibson assembly: the exonuclease contained in the Master Mix uncovered the Lox Recombination sites because they were too close to the overlapping ends.
Interlab Study:
Different strain for qPCR and Fluorescence test/ Modeling:
Because of the absence of experimental results, we can't model our enzymatic system.
CRE: - the yeast assembly did not work. pNP assay: - Two of our tests are less decisive because we didn't wait for a long time to see a real difference enter control and modified bacteria. - transformation of the construct very hard.
^
Page up
TPA toxicity
- Assessment of the toxicity
- Assessment of the toxicity
Operon assembly
- Succesful assembly of 2 of our 4 gene clusters (BBa_K1622000 and BBa_K1622001).
- Optimization of DNA sequences for E.coli.
Interlab Study
- Successful building of the 3 devices
- Characterization of the 3 devices using a Tecan micro-plate reader.
- Quantification of the number of Plasmids in each bacteria.
- Determination of the best Promoter
Here you can describe the results of your project and your future plans.
What should this page contain?
- Clearly and objectively describe the results of your work.
- Future plans for the project
- Considerations for replicating the experiments
Project Achievements
You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.
- A list of linked bullet points of the successful results during your project
- A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.
Inspiration
See how other teams presented their results.
Gibson assembly:
- BioBricks submitted to be BioBrick registry:
- Bba_K1622000: assembly of the Glycoaldehyde Dehydrogenase (BBa_K936011) and the Glycoaldehyde Reductase (BBa_K936023) in the plasmid PSB1C3 :
- Bba_K1622001: assembly of the NB-Esterase (BBa_K808030) and the TPA transporteur (BBa_K808007)
Interlab Study:
- Transformation in DH5-alpha of each promoter (BBa_J23101, BBa_J23106, BBa_J23117) and the controls GFP (BBa_I13405) and WT.
- qPCR of the transformed cells to determinate the plasmid copy number per strain.
- fluorescence test of the GFP expression and determination of the fluorescence per plasmid.
pNP assay:
- Insertion of the NB-Esterase (BBa_K808030) in the plasmid pDG011
- Transformation of BAP1 with the construct NB-Esterase (Bba_K808030) in the plasmid pDG011
- Isolation of three clones who have our construct NB-Esterase (Bba_K808030) in the plasmid pDG011.
- Demonstration of the degradation of the 4-NitroPhenyl Butyrate by our modified BAP1 with NB-Esterase in the pDG011 plasmid.
TPA :
- Test of the TPA toxicity thanks to a variation of the TPA concentration.
- Solubilization of TPA and amelioration of the method.
Problems:
Gibson assembly: the exonuclease contained in the Master Mix uncovered the Lox Recombination sites because they were too close to the overlapping ends.
Interlab Study:
Different strain for qPCR and Fluorescence test/ Modeling:
Because of the absence of experimental results, we can't model our enzymatic system.
CRE: - the yeast assembly did not work. pNP assay: - Two of our tests are less decisive because we didn't wait for a long time to see a real difference enter control and modified bacteria. - transformation of the construct very hard.
^
Page up
- Succesful assembly of 2 of our 4 gene clusters (BBa_K1622000 and BBa_K1622001).
- Optimization of DNA sequences for E.coli.
- Successful building of the 3 devices
- Characterization of the 3 devices using a Tecan micro-plate reader.
- Quantification of the number of Plasmids in each bacteria.
- Determination of the best Promoter
- Clearly and objectively describe the results of your work.
- Future plans for the project
- Considerations for replicating the experiments
- A list of linked bullet points of the successful results during your project
- A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.
Interlab Study
Here you can describe the results of your project and your future plans.
What should this page contain?
Project Achievements
You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.
Inspiration
See how other teams presented their results.
Gibson assembly:
- BioBricks submitted to be BioBrick registry:
- Bba_K1622000: assembly of the Glycoaldehyde Dehydrogenase (BBa_K936011) and the Glycoaldehyde Reductase (BBa_K936023) in the plasmid PSB1C3 :
- Bba_K1622001: assembly of the NB-Esterase (BBa_K808030) and the TPA transporteur (BBa_K808007)
Interlab Study:
- Transformation in DH5-alpha of each promoter (BBa_J23101, BBa_J23106, BBa_J23117) and the controls GFP (BBa_I13405) and WT.
- qPCR of the transformed cells to determinate the plasmid copy number per strain.
- fluorescence test of the GFP expression and determination of the fluorescence per plasmid.
pNP assay:
- Insertion of the NB-Esterase (BBa_K808030) in the plasmid pDG011
- Transformation of BAP1 with the construct NB-Esterase (Bba_K808030) in the plasmid pDG011
- Isolation of three clones who have our construct NB-Esterase (Bba_K808030) in the plasmid pDG011.
- Demonstration of the degradation of the 4-NitroPhenyl Butyrate by our modified BAP1 with NB-Esterase in the pDG011 plasmid.
TPA :
- Test of the TPA toxicity thanks to a variation of the TPA concentration.
- Solubilization of TPA and amelioration of the method.
Problems:
Gibson assembly: the exonuclease contained in the Master Mix uncovered the Lox Recombination sites because they were too close to the overlapping ends.
Interlab Study:
Different strain for qPCR and Fluorescence test/ Modeling:
Because of the absence of experimental results, we can't model our enzymatic system.
CRE: - the yeast assembly did not work. pNP assay: - Two of our tests are less decisive because we didn't wait for a long time to see a real difference enter control and modified bacteria. - transformation of the construct very hard.
^
Page up