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Team:Calgary/Notebook/Protocols/BenzylpenicillianAssay
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<h2>Reagents and Materials</h2> | <h2>Reagents and Materials</h2> | ||
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<li>96-well plate</li> | <li>96-well plate</li> | ||
<li>0.5% m/v Phenol Red</li> | <li>0.5% m/v Phenol Red</li> | ||
| - | <li> | + | <li>Penicillim G</li> |
<li>NaOH 1M</li> | <li>NaOH 1M</li> | ||
<li>Purified Beta-lactamase 1µg/µL</li> | <li>Purified Beta-lactamase 1µg/µL</li> | ||
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<ol> | <ol> | ||
<li>Add different amounts Beta-lactamase to the wells (we added 5µg to 25µg of purified TALE A-linker-beta-lactamase)</li> | <li>Add different amounts Beta-lactamase to the wells (we added 5µg to 25µg of purified TALE A-linker-beta-lactamase)</li> | ||
| - | <li>To a tube add 16.6mL of ddH<sub>2</sub>O (deionized water), 2mL of 0.5% m/v Phenol Red and 1.2g of | + | <li>To a tube add 16.6mL of ddH<sub>2</sub>O (deionized water), 2mL of 0.5% m/v Phenol Red and 1.2g of benzylpenicillin. Add the pH to 8.5 with NaOH 1M</li> |
<li>Add 100µL of the solution above to each well</li> | <li>Add 100µL of the solution above to each well</li> | ||
<li>Incubate at room temperature for 10 minutes</li> | <li>Incubate at room temperature for 10 minutes</li> | ||
Revision as of 00:52, 28 September 2013
Benzylpenicillin Assay
Benzylpenicillin Assay
Reagents and Materials
- 96-well plate
- 0.5% m/v Phenol Red
- Penicillim G
- NaOH 1M
- Purified Beta-lactamase 1µg/µL
- pH strip
Protocol
- Add different amounts Beta-lactamase to the wells (we added 5µg to 25µg of purified TALE A-linker-beta-lactamase)
- To a tube add 16.6mL of ddH2O (deionized water), 2mL of 0.5% m/v Phenol Red and 1.2g of benzylpenicillin. Add the pH to 8.5 with NaOH 1M
- Add 100µL of the solution above to each well
- Incubate at room temperature for 10 minutes
- Observe color change from red to yellow
- Measure pH with a pH strip (the pH is expected to drop)
