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| Line 6: |
Line 6: |
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| | <h3> Project Results</h3> | | <h3> Project Results</h3> |
| − | <hr>
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| − | <h5>11/09/15</h5>
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| − | <p>
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| − | Nanodrop results from plasmid prep<br>
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| − | OmcA (1) - 260/280= 1.87 260/230=2.10<br>
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| − | TOTAL= 114.5 ng/µl<br>
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| − | OmcA (2) - 260/280= 1.88 260/230=2.13<br>
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| − | TOTAL= 77.9 ng/µl<br>
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| − | CymA (1) - 260/280= 1.86 260/230=2.13<br>
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| − | TOTAL= 183.5ng/µl<br>
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| − | CymA (2) - 260/280= 1.88 260/230=2.06<br>
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| − | TOTAL= 107.1ng/µl<br>
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| − | MtrC (1) - 260/280= 1.88 260/230=2.15<br>
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| − | TOTAL= 166.7ng/µl<br>
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| − | MtrC (2) - 260/280= 1.89 260/230=2.18<br>
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| − | TOTAL= 91.1 ng/µl</p>
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| − | <hr>
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| − | <h5>17/09/15</h5><p>
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| − | 2 Gels of PCR products extracted yesterday morning (16/09/15) (tube 1, 2, 3, 4, 5, 6, 7, 8, MtrCAB). Gel was run for 40 mins at 100V. <br>
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| − | GEL 1</b><br>
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| − | <table border=="1" >
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| − | <tbody>
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| − | <tr>
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| − | <td valign="top" ><p><b>Lane 1</b></p></td>
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| − | <td valign="top" ><p><b>Lane 2</b></p></td>
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| − | <td valign="top" ><p><b>Lane 3</b></p></td>
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| − | <td valign="top" ><p><b>Lane 4</b></p></td>
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| − | <td valign="top" ><p><b>Lane 5</b></p></td>
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| − | <td valign="top" ><p><b>Lane 6</b></p></td>
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| − | <td valign="top" ><p><b>Lane 7</b></p></td>
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| − | <td valign="top" ><p><b>Lane 8</b></p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td>
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| − | <td valign="top" ><p><b>MtrCAB (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 1 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 2 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 3 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 4 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td>
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| − | <td valign="top" ><p><b>Empty</b></p></td>
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| − | </tr>
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| − |
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| − | </tbody>
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| − | </table>
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| − |
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| − |
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| − | <br><br>Both ladders and MtrCAB were the only lanes with results. <br>
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| − |
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| − | <br><br> <br> <br> <br> <br> <br>image here <br> <br> <br> <br> <br> <br>
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| − |
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| − |
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| − |
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| − |
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| − | <b>GEL 2 </b><br><table border="1">
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| − | <tbody>
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| − | <tr>
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| − | <td valign="top" ><p><b>Lane 1</b></p></td>
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| − | <td valign="top" ><p><b>Lane 2</b></p></td>
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| − | <td valign="top" ><p><b>Lane 3</b></p></td>
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| − | <td valign="top" ><p><b>Lane 4</b></p></td>
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| − | <td valign="top" ><p><b>Lane 5</b></p></td>
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| − | <td valign="top" ><p><b>Lane 6</b></p></td>
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| − | <td valign="top" ><p><b>Lane 7</b></p></td>
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| − | <td valign="top" ><p><b>Lane 8</b></p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 5 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 6 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 7 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>Tube 8 (2 µl + 8 µl)</b></p></td>
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| − | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td>
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| − | <td valign="top" ><p><b>Empty</b></p></td>
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| − | <td valign="top" ><p><b>Empty</b></p></td>
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| − | </tr>
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| − |
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| − | </tbody>
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| − | </table>
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| − |
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| − |
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| − | <br>
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| − | Lane 3 was the only lane without results.
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| − | <br> <br> <br> <br> <br> <br>image here <br> <br> <br> <br> <br> <br>
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| − |
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| − |
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| − | <br>
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| − | <b>Plasmid prep</b> <br>
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| − | 9 LB bottles were inoculated with amplicillin (100µg/ml) or chlorophenicol (34.5µg/ml) and plasmid pSB1C3 or pSB1A3 from yesterday’s transformation and left to incubate overnight (pSB1C3 =tube 2, 4, 5, 7 and MtrCAB; pSB1A3= tube 1, 3, 6, 8).<br><br>
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| − |
| |
| − | <table border="1">
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| − | <tbody>
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| − | <tr>
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| − | <td valign="top" ><p>Steps</p></td>
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| − | <td valign="top" ><p>Tube 1,3,6,8 (Amp)</p></td>
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| − | <td valign="top" ><p>Tube 2,5,7 (all Chlr) MtrCAB, </p></td>
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| − | <td valign="top" ><p>Tube 4 Chlr</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>1. Harvest</p></td>
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| − | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td>
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| − | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td>
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| − | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>2. Resuspend</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>3. Lyse</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>4. Precipitate</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>5. Bind</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>6. Wash (optional)</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>7. Wash and Ethanol Removal</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>8. Elute</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p>9. Recover</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | <td valign="top" ><p>yes</p></td>
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| − | </tr>
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| − |
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| − | </tbody>
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| − | </table>
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| − |
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| − | <br>
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| − |
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| − |
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| − | Tube 3 (Amp) - resuspended with 340 µl rather than 250 µl <br>
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| − | Tube 7 (Chlr) - 360 µl resuspension buffer added rather than 250 µl <br>
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| − | <b>Nanodrop results of plasmid prep </b><br>
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| − | <b>Amp 1</b>- 260/280= 1.98 260/230=2.12 <br>
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| − | TOTAL= 83.0ng/µl <br>
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| − | <b>Chlr 2</b>- 260/280= 1.89 260/230=2.11 <br>
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| − | TOTAL= 151.7ng/µl <br>
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| − | <b>Amp 3</b>- 260/280= 1.91 260/230=2.12 <br>
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| − | TOTAL= 173.5ng/µl <br>
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| − | <b>Chlr 4</b>- 260/280= 1.89 260/230=2.18 <br>
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| − | TOTAL= 92.1ng/µl <br>
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| − | <b>Chlr 5</b>- 260/280= 1.93 260/230=2.23 <br>
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| − | TOTAL= 87.8ng/µl <br>
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| − | <b>Amp 6</b>- 260/280= 1.92 260/230=2.15 <br>
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| − | TOTAL= 129.5ng/µl <br>
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| − | <b>Chlr 7</b>- 260/280= 1.91 260/230=2.21 <br>
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| − | TOTAL= 80.4ng/µl <br>
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| − | <b>Chlr 8</b>- 260/280= 1.94 260/230=2.11 <br>
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| − | TOTAL= 128.4ng/µl <br>
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| − | <b>MtrCAB</b>- 260/280= 1.96 260/230=2.28 <br>
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| − | TOTAL= 38.3ng/µl <br> <br>
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| − |
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| − | Below is the plate that the plasmid DNA that will be freeze dried <br>
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| − |
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| − |
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| − |
| |
| − |
| |
| − | <table border="1">
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| − | <tbody>
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| − | <tr>
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| − | <td valign="top" ><p><br />
| |
| − | </p></td>
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| − | <td valign="top" ><p><b>1</b></p></td>
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| − | <td valign="top" ><p><b>2</b></p></td>
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| − | <td valign="top" ><p><b>3</b></p></td>
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| − | <td valign="top" ><p><b>4</b></p></td>
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| − | </tr>
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| − |
| |
| − | <tr>
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| − | <td valign="top" ><p><b>A</b></p></td>
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| − | <td valign="top" ><p>1</p></td>
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| − | <td valign="top" ><p>9</p></td>
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| − | <td valign="top" ><p>17</p></td>
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| − | <td valign="top" ><p>25</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>B</b></p></td>
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| − | <td valign="top" ><p>2</p></td>
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| − | <td valign="top" ><p>10</p></td>
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| − | <td valign="top" ><p>18</p></td>
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| − | <td valign="top" ><p>26</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>C</b></p></td>
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| − | <td valign="top" ><p>3</p></td>
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| − | <td valign="top" ><p>11</p></td>
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| − | <td valign="top" ><p>19</p></td>
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| − | <td valign="top" ><p>27</p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>D</b></p></td>
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| − | <td valign="top" ><p>4</p></td>
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| − | <td valign="top" ><p>12</p></td>
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| − | <td valign="top" ><p>20</p></td>
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| − | <td valign="top" ><p><br />
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| − | </p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>E</b></p></td>
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| − | <td valign="top" ><p>5</p></td>
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| − | <td valign="top" ><p>13</p></td>
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| − | <td valign="top" ><p>21</p></td>
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| − | <td valign="top" ><p><br />
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| − | </p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>F</b></p></td>
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| − | <td valign="top" ><p>6</p></td>
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| − | <td valign="top" ><p>14</p></td>
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| − | <td valign="top" ><p>22</p></td>
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| − | <td valign="top" ><p><br />
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| − | </p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>G</b></p></td>
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| − | <td valign="top" ><p>7</p></td>
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| − | <td valign="top" ><p>15</p></td>
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| − | <td valign="top" ><p>23</p></td>
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| − | <td valign="top" ><p><br />
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| − | </p></td>
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| − | </tr>
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| − |
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| − | <tr>
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| − | <td valign="top" ><p><b>H</b></p></td>
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| − | <td valign="top" ><p>8</p></td>
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| − | <td valign="top" ><p>16</p></td>
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| − | <td valign="top" ><p>24</p></td>
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| − | <td valign="top" ><p><br />
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| − | </p></td>
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| − | </tr>
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| − |
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| − | </tbody>
| |
| − | </table>
| |
| − | <br>
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| − | <table><tbody><tr><td align="left">
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| − | 1. MtrA pSB1C3 <br>
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| − | 2. MtrB pSB1C3 <br>
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| − | 3. MtrC pSB1C3 <br>
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| − | 4. MtrC (1) <br>
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| − | 5. MtrC (2) <br>
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| − | 6. CymA (1) <br>
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| − | 7. CymA (2) </td><td align="left">
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| − | 8. OmcA <br>
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| − | 9. OmcA (1) <br>
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| − | 10. OmcA (2) <br>
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| − | 11. MtrA-His <br>
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| − | 12. MtrB-His <br>
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| − | 13. MtrC-His <br>
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| − | 14. CymA-His <br>
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| − | 15. OmcA-His <br>
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| − | 16- OmcA-CymA(1)</td><td>
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| − | 17. OmcA-CymA (2) <br>
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| − | 18. MtrCAB <br>
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| − | 19. Chlr 2 <br>
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| − | 20. Chlr 4 <br>
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| − | 21. Chlr 5 <br>
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| − | 22. Chlr 7</td></tr></tbody></table>
| |
| − |
| |
| − | </p>
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| − |
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| − |
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