Team:Newcastle/22 July 2010

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===Aim of this experiment===
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=Arginine Test=
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==Aim==
The aim of this experiment is to determine whether ''B. subtilis'' 168 is able to take up external arginine.
The aim of this experiment is to determine whether ''B. subtilis'' 168 is able to take up external arginine.
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===Materials Required===
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==Procedure==
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* Plate consisting of ''Bacillus subtilis'' 168 colonies.
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* Please refer to [[Team:Newcastle/Arginine test| Arginine test]]
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* Flame (streaking) Loop
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* LB media consisting arginine and ampicillin
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* Auto pipette
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* Bursen Burner
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* Universal Tube
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===Procedure===
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==Expected results==
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* Perform the experiment using aseptic technique.
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Arginine is an amino acid that is acidic. Therefore if ''B. subtilis'' 168 is able to take up arginine, it will cause a pH change in the media. This would result in an increase in pH.
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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.
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* Transfer 1 ml of the overnight culture to another universal tube containing 4 ml of the following media:
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#Control (1) - LB media
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#Control (2) - LB media with 10 mM of arginine
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#Control (3) - LB media plus ''B. subtilis'' 168
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#Test (1) - LB media with 10 mM of arginine plus ''B. subtilis'' 168
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#Test (2) - LB media with 10 mM of arginine plus ''B. subtilis'' 168
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* Incubate the culture at 37° C with shaking.
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* Record the pH at every 30 min interval.Use 20 ul of the culture and measure the pH using the pH measuring stick.
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===Inference===
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#Control (1) - No change in pH
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Cells will grow overnight at an exponential phase in the universal tube consisting media.
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#Control (2) - No change in pH
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#Control (3) - Increase in pH, however will be lower than test 1 and test 2.
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#Test (1) - Increase in pH
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#Test (2) - Increase in pH
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For actual results, please see [[Team:Newcastle/23_July_2010|23.07.10]] lab results.  
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==Conclusion==
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For experimental conclusion, please see 23.07.2010 lab results.
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{{Team:Newcastle/footer}}

Latest revision as of 21:47, 25 October 2010

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Contents

Arginine Test

Aim

The aim of this experiment is to determine whether B. subtilis 168 is able to take up external arginine.

Procedure

Expected results

Arginine is an amino acid that is acidic. Therefore if B. subtilis 168 is able to take up arginine, it will cause a pH change in the media. This would result in an increase in pH.

  1. Control (1) - No change in pH
  2. Control (2) - No change in pH
  3. Control (3) - Increase in pH, however will be lower than test 1 and test 2.
  4. Test (1) - Increase in pH
  5. Test (2) - Increase in pH

For actual results, please see 23.07.10 lab results.

Conclusion

For experimental conclusion, please see 23.07.2010 lab results.

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