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Team:Newcastle/22 July 2010
From 2010.igem.org
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* Transfer ''B. subtilis'' 168 colonies into universal tubes containing 5 ml of LB media and allowed to grow overnight at 37° C. | * 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: | * Transfer 1 ml of the overnight culture to another universal tube containing 4 ml of the following media: | ||
| - | # | + | #Control (1) - LB media |
| - | # | + | #Control (2) - LB media with 10 mM of arginine |
| - | #Test 1 - LB media with 10 mM of arginine plus | + | #Control (3) - LB media plus ''B. subtilis'' 168 |
| + | #Test (1) - LB media with 10 mM of arginine plus ''B. subtilis'' 168 | ||
| + | #Test (2) - LB media with 10 mM of arginine plus ''B. subtilis'' 168 | ||
* Incubate the culture at 37° C with shaking. | * 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. | + | * Record the pH at every 30 min interval.Use 20 ul of the culture and measure the pH using the pH measuring stick. |
===Inference=== | ===Inference=== | ||
Revision as of 15:43, 22 July 2010
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Contents |
Aim of this experiment
The aim of this experiment is to determine whether B. subtilis 168 is able to take up external arginine.
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:
- Control (1) - LB media
- Control (2) - LB media with 10 mM of arginine
- Control (3) - LB media plus B. subtilis 168
- Test (1) - LB media with 10 mM of arginine plus B. subtilis 168
- 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.
Inference
Cells will grow overnight at an exponential phase in the universal tube consisting media.
   
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