Team:Calgary/Notebook/Protocols/BenzylpenicillianAssay

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<div id="Banner"><h1>Benzylpenicillian Assay</h1></div>
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<div id="Banner"><h1>Benzylpenicillin Assay</h1></div>
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<h1>Benzylpenicillian Assay</h1>
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<h1>Benzylpenicillin Assay</h1>
<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>
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<li>Penicillium G</li>
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<li>Benzylpenicillin</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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<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>
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<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 Penicillium G. Add the pH to 8.5 with NaOH 1M</li>
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<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>

Latest revision as of 00:53, 28 September 2013

Benzylpenicillin Assay

Reagents and Materials

  • 96-well plate
  • 0.5% m/v Phenol Red
  • Benzylpenicillin
  • NaOH 1M
  • Purified Beta-lactamase 1µg/µL
  • pH strip

Protocol

  1. Add different amounts Beta-lactamase to the wells (we added 5µg to 25µg of purified TALE A-linker-beta-lactamase)
  2. 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
  3. Add 100µL of the solution above to each well
  4. Incubate at room temperature for 10 minutes
  5. Observe color change from red to yellow
  6. Measure pH with a pH strip (the pH is expected to drop)

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