Team:Warsaw/Calendar-Main/23 May 2008

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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#digest">Digestation> of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI</p>
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<h3>Rifampicin test #2</h3>
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<h4>Piotr</h4>
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<p>2. Digestion of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI</p>
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<p> Plating cultures from the previous day on rifampicin.</p>
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<p>3. <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</p>
 
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<p>4. Digest of pMPM-T5+AID with HindIII and SalI</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>
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<p>5. Digest of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI</p>
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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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<p>6. 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>
 
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<p>7. Ligation of purified products from p.1 and p.4</p>
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<p>8. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Electroporation</a> of E.coli TOP10 with ligation product</p>
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<p>9. Ligation of digested products from p.4 and p.5</p>
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<p>10. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Electroporation</a> of E.coli TOP10 with ligation product</p>
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<p>11. Transformants plating on LB + tetracycline</p>
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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).

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