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Team:Newcastle/Arginine test
From 2010.igem.org
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* Perform the experiment using aseptic technique. | * Perform the experiment using aseptic technique. | ||
* Transfer ''B. subtilis'' 168 colonies into universal tubes containing 5 ml of LB media and allowed to grow overnight at 37° C. | * Transfer ''B. subtilis'' 168 colonies into universal tubes containing 5 ml of LB media and allowed to grow overnight at 37° C. | ||
Revision as of 15:19, 28 July 2010
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Materials Required
- Plate consisting of Bacillus subtilis 168 colonies.
- Flame (streaking) Loop
- LB media consisting arginine and ampicillin
- Auto pipette
- Bursen Burner
- Universal Tube
Procedures
- Perform the experiment using aseptic technique.
- Transfer B. subtilis 168 colonies into universal tubes containing 5 ml of LB media and allowed to grow overnight at 37° C.
- Transfer 1 ml of the overnight culture to another universal tube containing 4 ml of the following media:
- Control (1) - LB media
- Control (2) - LB media with 10 mM of arginine
- Control (3) - LB media plus B. subtilis 168
- Test (1) - LB media with 10 mM of arginine plus B. subtilis 168
- Test (2) - LB media with 10 mM of arginine plus B. subtilis 168
- Incubate the culture at 37° C with shaking.
- Record the pH at every 30 min interval.Use 20 ul of the culture and measure the pH using the pH measuring stick.
   
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