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Team:Newcastle/29 June 2010
From 2010.igem.org
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29 June 2010
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Aims
The aim of this experiment is to prove that Bacillus subtilis 168 can take up arginine from the media into the cell.
Materials
- pH strips
- LB media
- arginine
- Universal Tubes
Protocol
- Set up and label the universal tubes as indicated:
- Negative controls- Media not containing Bacillus subtilis 168 (with/without arginine), LB without arginine innoculated with Bacillus subtilis 168
- Test- Media with arginine inoculated with Bacillus subtilis 168 cells
- Transfer 4ml of appropriate media into two universals
- Add 1ml of overnight culture to the appropriate tubes
- Incubate at 37°C at 180rpm
- Record the pH change of the media after every 1 hour by removing 20 µl of the media from both control and test tubes.
Result
| Hours | Negative control | Test |
|---|---|---|
| 0 | pH 7 | pH 7 |
| 1 | pH 7 | pH 7 |
| 2 | pH 7 | pH 7.5 |
| 3 | pH 7 | pH 7.5 |
| 4 | pH 7 | pH 8 |
| 5 | pH 7 | pH 8 |
Discussion
Arginine consists of a positive charge on it and thus is acidic in nature. As the cells take up the arginine from the media, the pH of the media increases. The tube labeled Negative control contains no cells and the pH of the media has remained constant for 5 hours which shows that the level of arginine in the media has remained constant, while the tube labeled Test which contains Bacillus subtilis 168 cells shows an increase in pH which shows that the level of arginine has decreased in the media.
Conclusion
It can be concluded that the Bacillus subtilis 168 cells are able to take up arginine from the media into the cells.
   
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