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Team:Newcastle/28 June 2010
From 2010.igem.org
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===Procedure=== | ===Procedure=== | ||
| - | * Unscrew the caps of | + | * Set up the universal tubes as indicated: |
| + | # Negative control- Media without ''Bacillus subtilis'' 168 | ||
| + | # Test- inoculated with ''Bacillus subtilis'' 168 | ||
| + | * Unscrew the caps of two 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. | ||
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* 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 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. | * 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 tubes on a shaker at 37° C. | + | * Put the two universal tubes on a shaker at 37° C. |
Revision as of 14:13, 21 July 2010
Aim of this experiment
The aim of this experiment was to prove that Bacillus subtilis 168 can take up arginine from the media into the cell.
Materials Required
- Plate consisting of Bacillus subtilis 168 colonies.
- Flaming (streaking) Loop
- LB media
- Auto pipette
- Bursen Burner
- Universal Tubes
Procedure
- Set up the universal tubes as indicated:
- Negative control- Media without Bacillus subtilis 168
- Test- inoculated with Bacillus subtilis 168
- Unscrew the caps of two 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 two universal tubes on a shaker at 37° C.
