Team:Calgary/Notebook/Protocols/BenzylpenicillianAssay2

From 2013.igem.org

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<h1>Protocols</h1>
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<h1>Benzylpenicillin Assay 2</h1>
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<h2>Reagents and Materials</h2>
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<ul>
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<li>96-well plate</li>
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<li>Bromothymol Blue pH Indicator</li>
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<li>60 mg/mL Benzylpenicillin</li>
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<li>Purified Beta-lactamase 1µg/µL</li>
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<li>Commercial Beta-lactamase 1µg/µL</li>
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<li>pH 8 Water</li>
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<h2>Protocol</h2>
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<li>Dilute beta-lactamase to the appropriate concentrations in 100 µL of pH 8 water. Add this to the wells.</li>
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<li>Add an additional 50 µL of pH 8 water to the wells.</li>
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<li>Add 10 µL of bromothymol blue pH indicator to these wells.</li>
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<li>Add 100 µL of 60 mg/mL benzylpenicillin to the wells.</li>
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<li>Measure the change from blue to yellow using a spectrophotometer at an absorbance wavelength of 616 nm.</li>
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Revision as of 00:52, 29 October 2013

Benzylpenicillin Assay 2

Reagents and Materials

  • 96-well plate
  • Bromothymol Blue pH Indicator
  • 60 mg/mL Benzylpenicillin
  • Purified Beta-lactamase 1µg/µL
  • Commercial Beta-lactamase 1µg/µL
  • pH 8 Water

Protocol

  1. Dilute beta-lactamase to the appropriate concentrations in 100 µL of pH 8 water. Add this to the wells.
  2. Add an additional 50 µL of pH 8 water to the wells.
  3. Add 10 µL of bromothymol blue pH indicator to these wells.
  4. Add 100 µL of 60 mg/mL benzylpenicillin to the wells.
  5. Measure the change from blue to yellow using a spectrophotometer at an absorbance wavelength of 616 nm.

Retrieved from "http://2013.igem.org/Team:Calgary/Notebook/Protocols/BenzylpenicillianAssay2"