Team:Newcastle/22 July 2010

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(New page: ===Chromosomal DNA Extraction of ''B. Subtilis'' Strain 168 and 3610=== Aim: Materials: LB Broth, ''B. Subtilis'' Strain 168 colonies and ''B. Subtilis'' Strain 3610 colonies, Procedur...)
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===Chromosomal DNA Extraction of ''B. Subtilis'' Strain 168 and 3610===
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{{Team:Newcastle/mainbanner}}
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Aim:
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===Aim of this experiment===
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The aim of this experiment is to determine whether ''B. subtilis'' 168 is able to take up external arginine.
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Materials: LB Broth, ''B. Subtilis'' Strain 168 colonies and ''B. Subtilis'' Strain 3610 colonies,
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===Materials Required===
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* Plate consisting of ''Bacillus subtilis'' 168 colonies.
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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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Procedures:
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===Procedure===
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* Perform the experiment using aseptic technique.
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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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#Negative control (1) - LB media
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#Negative control (2) - LB media with 10 mM of arginine
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#Test 1 - LB media with 10 mM of arginine plus 
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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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# 5ml of LB Broth is pipetted into a universal tube, two tubes of each strain were prepared.
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===Inference===
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# Loop is heated to pick colonies from agar plates.
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Cells will grow overnight at an exponential phase in the universal tube consisting media.
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# All tubes are left overnight in the 37°C room.
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Revision as of 15:42, 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:
  1. Negative control (1) - LB media
  2. Negative control (2) - LB media with 10 mM of arginine
  3. Test 1 - LB media with 10 mM of arginine plus
  • 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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