Team:Newcastle/Urease test
From 2010.igem.org
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{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
- | === | + | =Urease test= |
+ | |||
+ | ==Materials required== | ||
*Pipettes | *Pipettes | ||
*Universal tubes | *Universal tubes | ||
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*Christensen's Urea Agar | *Christensen's Urea Agar | ||
**Make up 1 liter of the agar mixture containing: | **Make up 1 liter of the agar mixture containing: | ||
- | *# Gelatine peptone 1. | + | *# Gelatine peptone 1.0 g |
- | *# Potassium dihydrogen phosphate 2. | + | *# Potassium dihydrogen phosphate 2.0 g |
- | *# Sodium chloride 5. | + | *# Sodium chloride 5.0 g |
- | *# Agar | + | *# Agar 20 g |
- | **Top up to 1 liter and apply gentle heating to dissolve. Sterilize at 115°C for at least | + | **Top up to 1 liter and apply gentle heating to dissolve. Sterilize at 115°C for at least 20 min. |
**Add the following to the molten base: | **Add the following to the molten base: | ||
- | *# D(+)-Glucose 1. | + | *# D(+)-Glucose 1.0 g |
- | *# Phenol red, 0.2% | + | *# Phenol red, 0.2% 6 ml |
**Note: Ensure that the work was done using aseptic technique. | **Note: Ensure that the work was done using aseptic technique. | ||
- | **Transfer | + | **Transfer 10 ml of the molten base to universal tube and allow the agar to set in a slanted position. |
**Store the harden agar in the fridge. | **Store the harden agar in the fridge. | ||
- | == | + | ==Procedures== |
# Perform the experiment using aseptic technique. | # Perform the experiment using aseptic technique. | ||
# Pick up single colony of ''B. subtilis'' 168 and streaking it onto the slanted agar tube. | # Pick up single colony of ''B. subtilis'' 168 and streaking it onto the slanted agar tube. |
Revision as of 15:21, 28 July 2010
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Urease test
Materials required
- Pipettes
- Universal tubes
- Streaking loop
- Christensen's Urea Agar
- Make up 1 liter of the agar mixture containing:
- Gelatine peptone 1.0 g
- Potassium dihydrogen phosphate 2.0 g
- Sodium chloride 5.0 g
- Agar 20 g
- Top up to 1 liter and apply gentle heating to dissolve. Sterilize at 115°C for at least 20 min.
- Add the following to the molten base:
- D(+)-Glucose 1.0 g
- Phenol red, 0.2% 6 ml
- Note: Ensure that the work was done using aseptic technique.
- Transfer 10 ml of the molten base to universal tube and allow the agar to set in a slanted position.
- Store the harden agar in the fridge.
Procedures
- 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