Team:Warsaw/Calendar-Main/23 May 2008

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<h4>Michał K.:</h4>
 
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of PCR product for transcription fusion (T7 RNA-polymerase) with HindIII and SalI (2x Tango buffer).</li>
 
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of PCR product for translation fusion (AID+T7 RNA-polymerase) with NcoI and XbaI (1x Tango buffer).</li>
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<h3>Rifampicin test #2</h3>
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<h4>Piotr</h4>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested products from p.1 and p.2.</li>
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<p> Plating cultures from the previous day on rifampicin.</p>
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<li>  <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).</li>
 
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<li> <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-omega>pMPM-T5</a> with NcoI and XbaI (1x Tango buffer).</li>
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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>
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<li> Gel-electrophresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands (p.4 - 4850 bp, p.5 - 4250 bp).</li>
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<p> Further inoculation of separate transformant colonies for transcriptional and translational fusions on liquid LB + tetracycline (as there were no positive colonies in previous assay).</p>
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<li>  <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of purified products from p.1 and p.4.</li>
 
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Electroporation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  with ligation product.</li>
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<li>  <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of digested products from p.4 and p.5.</li>
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<li>  <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Electroporation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  with ligation product.</li>
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<li>  Transformants plating on LB + tetracycline.</li></ol>
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http://openwetware.org/images/thumb/6/61/UW_A%2BT.jpg/350px-UW_A%2BT.jpg http://openwetware.org/images/thumb/c/c7/UW_AT.jpg/350px-UW_AT.jpg
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Latest revision as of 14:08, 26 October 2008

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Rifampicin test #2

Piotr

Plating cultures from the previous day on rifampicin.

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

Michał K.

Further inoculation of separate transformant colonies for transcriptional and translational fusions on liquid LB + tetracycline (as there were no positive colonies in previous assay).