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Team:Newcastle/29 June 2010
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Shethharsh08 (Talk | contribs) (New page: ===Aim of this experiment=== The aim of this experiment is to prove that ''Bacillus subtilis'' 168 can take up arginine from the media into the cell. ===Materials Required=== * pH strips ...) |
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===Procedure=== | ===Procedure=== | ||
| - | * | + | * Set up and label the universal tubes as indicated: |
| - | * Unscrew the caps of the universal | + | # Negative control- Media not containing ''Bacillus subtilis'' 168 |
| + | # Test- Media inoculated with ''Bacillus subtilis'' 168 cells. | ||
| + | * Unscrew the caps of the universal tubes and after unscrewing, flame the mouth of the tube. | ||
* With the help of an auto pipette, pipette out 5ml of LB broth from the stock solution and transfer it into universal tubes. Do this process three times so as to transfer 5ml of LB media in all the three tubes. | * With the help of an auto pipette, pipette out 5ml of LB broth from the stock solution and transfer it into universal tubes. Do this process three times so as to transfer 5ml of LB media in all the three tubes. | ||
* Flame the flaming loop with the help of a burner till the loop gets red in colour. | * Flame the flaming loop with the help of a burner till the loop gets red in colour. | ||
Revision as of 14:25, 21 July 2010
Contents |
Aim of this experiment
The aim of this experiment is to prove that Bacillus subtilis 168 can take up arginine from the media into the cell.
Materials Required
- pH strips
- LB media consisting arginine and ampicillin
- Auto pipette
- Gilson pipette
- Autoclaved tips for gilson pipette
- Burnsen Burner
- Universal Tube
Procedure
- Set up and label the universal tubes as indicated:
- Negative control- Media not containing Bacillus subtilis 168
- Test- Media inoculated with Bacillus subtilis 168 cells.
- Unscrew the caps of the universal tubes and after unscrewing, flame the mouth of the tube.
- With the help of an auto pipette, pipette out 5ml of LB broth from the stock solution and transfer it into universal tubes. Do this process three times so as to transfer 5ml of LB media in all the three tubes.
- Flame the flaming loop with the help of a burner till 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 flaming loop with the help of a burner till the loop gets red in colour so as to kill the cells present on it.
- 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.
