Team:Newcastle/Urease test
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===Materials=== | ===Materials=== | ||
*Pipettes | *Pipettes |
Revision as of 14:48, 28 July 2010
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Materials
- Pipettes
- Universal tubes
- Streaking loop
- Christensen's Urea Agar
- Make up 1 liter of the agar mixture containing:
- Gelatine peptone 1.0g
- Potassium dihydrogen phosphate 2.0g
- Sodium chloride 5.0g
- Agar 20g
- Top up to 1 liter and apply gentle heating to dissolve. Sterilize at 115°C for at least 20min.
- Add the following to the molten base:
- D(+)-Glucose 1.0g
- Phenol red, 0.2% 6ml
- Note: Ensure that the work was done using aseptic technique.
- Transfer 10ml of the molten base to universal tube and allow the agar to set in a slanted position.
- Store the harden agar in the fridge.
Protocol
- Perform the experiment using aseptic technique.
- Pick up single colony of B. subtilis 168 and streaking it onto the slanted agar tube.
- Loosen up the cap to allow air exchange between the interior of the tube and the external environment.
- Incubate the tubes overnight at 37°C.
- Set up the tubes as indicated:
- Negative control - Without B. subtilis 168
- Test 1 (Duplicate) - Inoculated with B. subtilis 168
- Test 2 (Duplicate) - Inoculated with B. subtilis 168