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- | <div id="Banner"><h1>Ampicillin Survival Assay 1</h1></div> | + | <div id="Banner"><h1>Ampicillin Survival Assay 2</h1></div> |
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| {{Team:Calgary/ContentPage}} | | {{Team:Calgary/ContentPage}} |
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- | <section id="Content"> | + | <section id="Content"> |
- | <h1>Ampicillin Survival Assay 1</h1> | + | |
| + | <h1>Ampicillin Survival Assay 2</h1> |
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- | <p class="noIndent">We used this protocol to verify if, after protein purification, beta-lactamase was functional when linked to one of our TALEs.</p> | + | <p>Insert Text Here</p> |
- | <h2>Reagents and Materials</h2>
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- | <ul>
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- | <li>LB Broth</li>
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- | <li>Ampicillin (100mg/mL)</li>
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- | <li>Chloramphenicol (30mg/mL)</li>
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- | <li>Falcon tubes</li>
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- | <li>Bacteria resistant to chloramphenicol but not ampicillin</li>
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- | <li>Bacteria resistant to both ampicillin and chloramphenicol</li>
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- | <li>Purified beta-lactamase protein</li>
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- | </ul>
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- | <h2>Protocol</h2>
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- | <ol>
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- | <li>To each Falcon tube, add 5mL of LB Broth, 5µL of ampicillin and 3µL of chloramphenicol</li>
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- | <li>Add your purified proteins in multiple tubes with different amounts of proteins; for construct we added the following: 20µg, 15µg, 10µg, 5µg, 1µg, 0.1µg and 0.01µg</li>
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- | <li>Incubate the tubes for 1h at 37°C</li>
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- | <li>For the positive control, inoculate a tube that doesn’t have protein on it with a colony of your bacteria resistant to both antibiotics</li>
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- | <li>Add colonies of your bacteria resistant to chloramphenicol but not ampicillin to the other tubes</li>
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- | <li>Grow the cells overnight</li>
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- | </ol>
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| </section> | | </section> |
| </html> | | </html> |