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Team:Newcastle/2 July 2010
From 2010.igem.org
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{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
| + | ==Urease Test== | ||
===Aims=== | ===Aims=== | ||
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# Test 2 (Duplicate) - Pink-red color | # Test 2 (Duplicate) - Pink-red color | ||
| - | === | + | ===Discussion=== |
''B. subtilis'' 168 is able to produce urease, therefore, urea which is the substrate and can be found in the agar was degraded, which results in ammonia building. The ammonia makes the media alkaline and therefore the indicator phenol red will change from orange color to pink-red color. | ''B. subtilis'' 168 is able to produce urease, therefore, urea which is the substrate and can be found in the agar was degraded, which results in ammonia building. The ammonia makes the media alkaline and therefore the indicator phenol red will change from orange color to pink-red color. | ||
| + | ===Conclusion=== | ||
The negative control did not turn pink-red color, therefore indicating that no contamination had occurred. | The negative control did not turn pink-red color, therefore indicating that no contamination had occurred. | ||
| + | |||
| + | ==LacI BioBrick Construction== | ||
| + | |||
| + | ===Aims=== | ||
| + | *To use PCR to extract ''lacI'' (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP). | ||
| + | |||
| + | ===Materials=== | ||
| + | *Competent ''E. coli'' (DH5alpha strain) | ||
| + | *pMutin4 plasmid | ||
| + | *pSB1AT3 plasmid | ||
| + | |||
| + | ===Protocol=== | ||
| + | * [[TeamNewcastleTransformation of E. coli|Transformation of ''E. coli'']] DH5alpha with pMutin4 and pSB1AT3 separately. | ||
| + | |||
| + | ===Inference=== | ||
| + | *''Ecoli'' DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells. | ||
| + | |||
| + | |||
| + | ==Competent ''E. coli'' Production== | ||
| + | |||
| + | ===Aims=== | ||
| + | *To make a stock of competent ''E. coli'' (DH5alpha strain) ready for transformation. | ||
| + | |||
| + | ===Materials=== | ||
| + | *''E. coli'' DH5alpha strain | ||
| + | |||
| + | ===Protocol=== | ||
| + | * Set up [[Team:Newcastle/Growing a liquid culture| liquid culture]] of ''E. coli'' DH5alpha | ||
| + | |||
| + | ===Inference=== | ||
| + | *Grow up a stock of ''E.coli'' DH5alpha strain ready to be made competent | ||
Revision as of 10:19, 27 July 2010
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Contents |
Urease Test
Aims
The aim of this experiment was to determine if Bacillus subtilis 168 is able to produce urease and degrade urea.
Urease test
The experiment was performed on 01.07.10. For the experimental protocol, please see 01.07.10 lab notebook.
Results
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Figure 1: Urease test results
- Negative control - No color change (Orange color)
- Test 1 (Duplicate) - Pink-red color
- Test 2 (Duplicate) - Pink-red color
Discussion
B. subtilis 168 is able to produce urease, therefore, urea which is the substrate and can be found in the agar was degraded, which results in ammonia building. The ammonia makes the media alkaline and therefore the indicator phenol red will change from orange color to pink-red color.
Conclusion
The negative control did not turn pink-red color, therefore indicating that no contamination had occurred.
LacI BioBrick Construction
Aims
- To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
Materials
- Competent E. coli (DH5alpha strain)
- pMutin4 plasmid
- pSB1AT3 plasmid
Protocol
- Transformation of E. coli DH5alpha with pMutin4 and pSB1AT3 separately.
Inference
- Ecoli DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells.
Competent E. coli Production
Aims
- To make a stock of competent E. coli (DH5alpha strain) ready for transformation.
Materials
- E. coli DH5alpha strain
Protocol
- Set up liquid culture of E. coli DH5alpha
Inference
- Grow up a stock of E.coli DH5alpha strain ready to be made competent
- Produce competent E. coli (DH5alpha strain)
   
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