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Team:Calgary/Notebook/Protocols/BenzylpenicillianAssay2
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<li>Bromothymol Blue pH Indicator</li> | <li>Bromothymol Blue pH Indicator</li> | ||
<li>60 mg/mL Benzylpenicillin</li> | <li>60 mg/mL Benzylpenicillin</li> | ||
| - | <li>Purified | + | <li>Purified β-lactamase 1µg/µL</li> |
| - | <li>Commercial | + | <li>Commercial β-lactamase 1µg/µL</li> |
<li>pH 8 Water</li> | <li>pH 8 Water</li> | ||
</ul> | </ul> | ||
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<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
| - | <li>Dilute beta-lactamase to the appropriate concentrations in 100 µL of pH 8 water. Add this to the wells.</li> | + | <li>Dilute β-lactamase to the appropriate concentrations in 100 µL of pH 8 water. Add this to the wells.</li> |
<li>Add an additional 50 µL of pH 8 water to the wells.</li> | <li>Add an additional 50 µL of pH 8 water to the wells.</li> | ||
<li>Add 10 µL of bromothymol blue pH indicator to these wells.</li> | <li>Add 10 µL of bromothymol blue pH indicator to these wells.</li> | ||
Latest revision as of 01:44, 29 October 2013
Benzylpenicillin Assay 2
Reagents and Materials
- 96-well plate
- Bromothymol Blue pH Indicator
- 60 mg/mL Benzylpenicillin
- Purified β-lactamase 1µg/µL
- Commercial β-lactamase 1µg/µL
- pH 8 Water
Protocol
- Dilute β-lactamase to the appropriate concentrations in 100 µL of pH 8 water. Add this to the wells.
- Add an additional 50 µL of pH 8 water to the wells.
- Add 10 µL of bromothymol blue pH indicator to these wells.
- Add 100 µL of 60 mg/mL benzylpenicillin to the wells.
- Measure the change from blue to yellow using a spectrophotometer at an absorbance wavelength of 616 nm.
