Team:Warsaw/Calendar-Main/24 May 2008

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<html><h3>Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7</h3>
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<html>
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<h4>Michał K.:</h4>
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<h3> Rifampicin test #2</h3>
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<h4>Piotr</h4>
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<table id="result">
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<tr><th>Strain</th><th>Inducer / repressor</th><th>Number of colonies on rifampicin</th></tr>
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<tr><th><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a></th><td>none</td><td>19</td></tr>
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<tr><th><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a></th><td>none</td><td>10</td></tr>
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<tr><th><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a></th><td>0.1% Arabinose</td><td>154</td></tr>
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<tr><th><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a></th><td>none</td><td>11</td></tr>
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<tr><th><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a></th><td>0.5 mM IPTG</td><td>59</td></tr>
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</table>
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<br/>
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<p>Conclusion: AID seems to work!</p>
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<h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a></h3>
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<h4>Michał K.</h4>
<p><ol>
<p><ol>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical pMPM-T5 with transcription fusion from other colonies.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a> from cultures.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical pMPM-T5 with translation fusion from other colonies.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a> from cultures.</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of pMPM-T5 with transcription fusion with EcoRI (EcoRI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a>with EcoRI (EcoRI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of pMPM-T5 with translation fusion with BamHI (BamHI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a> with BamHI (BamHI buffer).</li>
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<li> Gel electrophoresis (still no proper colonies found).</li></html>
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<li> Gel electrophoresis (still no proper colonies found).</li>
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</ol></p>
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</html>

Latest revision as of 14:09, 26 October 2008

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Rifampicin test #2

Piotr

StrainInducer / repressorNumber of colonies on rifampicin
Top10none19
Top10 pMPM-T5-AIDnone10
Top10 pMPM-T5-AID0.1% Arabinose154
Top10 pTrc99a-AIDnone11
Top10 pTrc99a-AID0.5 mM IPTG59

Conclusion: AID seems to work!

Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

  1. Isolation of hypothetical pMPM-T5 with transcriptional fusion from cultures.
  2. Isolation of hypothetical pMPM-T5 with translational fusion from cultures.
  3. Control digest of pMPM-T5 with transcriptional fusionwith EcoRI (EcoRI buffer).
  4. Control digest of pMPM-T5 with translational fusion with BamHI (BamHI buffer).
  5. Gel electrophoresis (still no proper colonies found).

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