From 2010.igem.org
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Arginine Test
Materials Required
- Plate containing Bacillus subtilis 168 colonies.
- Flame (streaking) loop
- LB media consisting of arginine and ampicillin
- Auto pipette
- Bursen Burner
- Universal Tube
- pH measuring stick
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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