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:

1. Control (1) - LB media
2. Control (2) - LB media with 10 mM of arginine
3. Control (3) - LB media plus B. subtilis 168
4. Test (1) - LB media with 10 mM of arginine plus B. subtilis 168
5. 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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