Team:Newcastle/Arginine test

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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

Procedure

  • 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.