Team:Newcastle/2 July 2010

Urease Test

Aims

The aim of this experiment was to determine if Bacillus subtilis 168 is able to produce urease and degrade urea.

Protocol

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

1. Negative control - No color change (Orange color)
2. Test 1 (Duplicate) - Pink-red color
3. 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)