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| - | <h2><b>V07_04 </b></h2><br>
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| - | <b> Repetition of the amplification of <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> </i></i></b><br>
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| - | <ul>
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| - | <li>Experiment: <br>
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| - | The genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Pfu-Polymerase according to the following protocol. Additionally, a negative control as well as a positive control were prepared this time. The success of the PCR was subsequently investigated preparing a 1% agarose gel.<br></li>
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| - | <li>Observations & Results: <br>
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| - | The did not function again for no bands were visible on the gel; only the marker was clearly discernible. However, since also the positive control did not feature the slightest band, we assume that something might be wrong with our polymerase.</li>
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| - | </table>
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| - | <h2><b>V07_05 </b></h2><br>
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| - | <b>V07_05_1 Repetition of the amplification of the genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i></b><br>
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| - | <ul>
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| - | <li>Experiment: <br>
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| - | To give it a last try the <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Pfu-Polymerase according to the following protocol one last time. Here again, a 1% agarose gel was prepared to investigate the PCRs outcome.</li><br>
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| - | <li>Observations & Results: <br>
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| - | The gel of the PCR delivered the same picture as last time. No band could be observed except for the marker. Even the positive control does not work. Thus, we decided to buy a new polymerase.</li>
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| - | </ul>
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| - | <br>
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| - | <b>V07_05_2 Repetition of the preparative double digestion of 20G-<i>flhDC</i>, 20I-<i>flhDC</i>, 18K-<i>flhDC</i>, and 18C-<i>flhDC</i></b><br>
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| - | <ul>
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| - | <li>Experiment: <br>
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| - | The test digestion of the <i>flhDC</i>-promoter constructs EcoRI and PstI were applied according to the protocol. The restriction products were subsequently separated on a gel in order to verify the digestions success and purify the desired fragments.<br>
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| - | <li>Observations & Results: <br>
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| - | The digestion was successful. Bands of the expected size could be observed and cut out.</li>
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| - | </ul><br>
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| - | </td></tr>
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| - | </table>
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| - | <table cellpadding="20 px" border="1" bordercolor="black" valign="top">
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| - | <h2><b>V07_06 </b></h2><br>
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| - | <b> Repetition of the amplification of the <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> using the new polymerase </b><br>
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| - | <ul>
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| - | <li>Experiment: <br>
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| - | The genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Phusion-Polymerase according to the following protocol. We also prepared a negative control as well as a positive control. The success of the PCR was subsequently investigated preparing a 1% analytical agarose gel.<br></li>
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| - | <li>Observations & Results: <br>
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| - | Finally,the PCR was successful! We were able to receive bands of the expected size for each construct. Obviously, the old Pfu-polymerase was not functional anymore and had caused the PCRs failure. </li>
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| - | <br></td></tr>
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| | </table> | | </table> |
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