Team:Warsaw/Calendar-Main/27 May 2008

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<html><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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<p>Michał K.:</p>
 
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<p>1. <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.</p>
 
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<p>2. <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.</p>
 
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<p>3. 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 and XhoI (2x Tango buffer).</p>
 
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<p>4. 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).</p>
 
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<p>5. 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 EcoRI and XbaI (2x Tango buffer).</p>
 
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<p>6. Choice of proper clones and preparation for sequencing.</p>
 
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<h3>Here we found the clone with translational fusion on pMPM. These are different isolations in each lane. We have been working with the AT6 ever since. <br>
 
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Digestion was conducted with BamHI</h3>
 
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<center>https://static.igem.org/mediawiki/2008/6/62/Trawienie_06_05_2008.jpg</center>
 
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<h4>Michał K.</h4>
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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 assays).
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</p> </html>

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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 assays).