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Team:Newcastle/29 June 2010
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* Set up and label the universal tubes as indicated: | * Set up and label the universal tubes as indicated: | ||
# Negative control- Media not containing ''Bacillus subtilis'' 168 | # Negative control- Media not containing ''Bacillus subtilis'' 168 | ||
| - | # Test- Media inoculated with ''Bacillus subtilis'' 168 cells | + | # Test- Media inoculated with ''Bacillus subtilis'' 168 cells |
* Unscrew the caps of the universal tubes and after unscrewing, flame the mouth of the tube. | * 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 | + | * With the help of an auto pipette, pipette out 4ml of LB broth from the stock solution and transfer it into the two universal tubes mentioned above. |
| - | * | + | * Remove the universal tube consisting of an overnight culture of growing ''Bacillus suntilis'' 168 cells and unscrew the cap and flame the mouth of the tube. |
| - | * | + | * Set the Gilson pipette to 1000 µl and attach a autoclaved tip to it. |
| - | * | + | * Suck 1000 µl out of the tube consisting of overnight culture and put it in the universal tube consisting 4ml of LB media. Do this step twice so as to put 1000 µl in both the control and the test tubes. |
* 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. | ||
Revision as of 14:32, 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 4ml of LB broth from the stock solution and transfer it into the two universal tubes mentioned above.
- Remove the universal tube consisting of an overnight culture of growing Bacillus suntilis 168 cells and unscrew the cap and flame the mouth of the tube.
- Set the Gilson pipette to 1000 µl and attach a autoclaved tip to it.
- Suck 1000 µl out of the tube consisting of overnight culture and put it in the universal tube consisting 4ml of LB media. Do this step twice so as to put 1000 µl in both the control and the test tubes.
- 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.
