Team:Newcastle/28 June 2010

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28 June 2010

Contents

Aims

The aim of this experiment is to prove that Bacillus subtilis 168 can take up arginine from the media into the cell.

Materials

  • Bacillus subtilis 168
  • LB media
  • arginine
  • Universal Tube

Protocol

  • Label the universal tube as "Overnight Culture".
  • Unscrew the caps of the universal tube and after unscrewing, flame the mouth of the tube.
  • With the help of an automatic pipette, pipette out 5ml of LB broth from the stock solution and transfer it into universal tubes.
  • Flame the flaming loop with the help of a burner until the loop gets red in colour.
  • Allow it to cool by holding the loop for approximately 10 seconds.
  • Use the loop to pick up a colony from an agar plate consisting of Bacillus subtilis 168 colonies and transfer the loop into the universal tube consisting the LB media.
  • Flame the mouth of the universal tubes and screw the cap (do not over tight the cap so as to allow oxygen to enter the tube)and shake gently so as to suspend the cells equally.
  • Flame the flame loop with the help of a burner until the loop turns red.
  • Put the universal tube on a shaker at 37° C and leave it overnight for the cells to grow.

Inference

Cells will grow overnight at an exponential phase in the universal tube consisting media.


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