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Team:Warsaw/Calendar-Main/3 June 2008
From 2008.igem.org
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<h3>Preparation of T7 constructs<br>Piotr</h3> | <h3>Preparation of T7 constructs<br>Piotr</h3> | ||
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<li>DNA (PCR products from previous day) gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of DNA from proper bands. </li> | <li>DNA (PCR products from previous day) gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of DNA from proper bands. </li> | ||
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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 reaction products.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of reaction products.</li> | ||
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion).</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion).</li> | ||
| - | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols# | + | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</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> |
<li>Plating transformants on LB+tetracycline.</li> | <li>Plating transformants on LB+tetracycline.</li> | ||
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of pMPM-T5 + transcription fusion with EcoRHI and HindIII.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of pMPM-T5 + transcription fusion with EcoRHI and HindIII.</li> | ||
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Isolation of DNA from proper band.</a></li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Isolation of DNA from proper band.</a></li> | ||
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of product p.11 (pMPM-T5 + T7 RNA-polymerase).</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of product p.11 (pMPM-T5 + T7 RNA-polymerase).</li> | ||
| - | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols# | + | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</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> |
<li>Plating transformants on LB + tetracycline.</li> | <li>Plating transformants on LB + tetracycline.</li> | ||
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<img src="https://static.igem.org/mediawiki/2008/a/a4/UW_AAT.jpg" width="500"/> | <img src="https://static.igem.org/mediawiki/2008/a/a4/UW_AAT.jpg" width="500"/> | ||
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Preparation of T7 constructs
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