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Team:Newcastle/1 July 2010
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| - | === | + | '''1 July 2010''' |
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| + | =Urease Test= | ||
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| + | ==Aims== | ||
The aim of this experiment was to determine if ''Bacillus subtilis'' 168 is able to produce urease and degrade urea. | The aim of this experiment was to determine if ''Bacillus subtilis'' 168 is able to produce urease and degrade urea. | ||
| - | + | ==Procedures== | |
* Please refer to [[Team:Newcastle/Urease test| urease test]] | * Please refer to [[Team:Newcastle/Urease test| urease test]] | ||
| - | + | ==Inference== | |
Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms. | Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms. | ||
The addition of gelatine peptone, dextrose and reduced content of buffer supports a luxuriant growth at an early stage. Urea is the substrate and can be degraded by certain organisms, 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. | The addition of gelatine peptone, dextrose and reduced content of buffer supports a luxuriant growth at an early stage. Urea is the substrate and can be degraded by certain organisms, 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. | ||
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For actual results, please see [[Team:Newcastle/2 July 2010#Results|02.07.10 lab notebook.]] | For actual results, please see [[Team:Newcastle/2 July 2010#Results|02.07.10 lab notebook.]] | ||
| - | + | =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). | *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) | *Competent ''E. coli'' (DH5alpha strain) | ||
*pMutin4 plasmid | *pMutin4 plasmid | ||
*pSB1AT3 plasmid | *pSB1AT3 plasmid | ||
| - | + | ==Protocol== | |
* [[TeamNewcastleTransformation of E. coli|Transformation of ''E. coli'']] DH5alpha with pMutin4 and pSB1AT3 separately. | * [[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. | *''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. | *To make a stock of competent ''E. coli'' (DH5alpha strain) ready for transformation. | ||
| - | + | ==Materials== | |
*''E. coli'' DH5alpha strain | *''E. coli'' DH5alpha strain | ||
| - | + | ==Protocol== | |
* Set up [[Team:Newcastle/Growing a liquid culture| liquid culture]] of ''E. coli'' DH5alpha | * 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 | *Grow up a stock of ''E.coli'' DH5alpha strain ready to be made competent | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 10:46, 11 August 2010
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1 July 2010
Contents |
Urease Test
Aims
The aim of this experiment was to determine if Bacillus subtilis 168 is able to produce urease and degrade urea.
Procedures
- Please refer to urease test
Inference
Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms. The addition of gelatine peptone, dextrose and reduced content of buffer supports a luxuriant growth at an early stage. Urea is the substrate and can be degraded by certain organisms, 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.
- Negative control - No color change (Orange color)
- Test 1 (Duplicate) - Pink-red color
- Test 2 (Duplicate) - Pink-red color
For actual results, please see 02.07.10 lab notebook.
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
   
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