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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">Digestion</a> of PCR product for transcription fusion (T7 RNA-polymerase) with HindIII and SalI (2x Tango buffer).</p> | + | <h3>Rifampicin test #2</h3> |
| | + | <h4>Piotr</h4> |
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| - | <p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestion</a> of PCR product for translation fusion (AID+T7 RNA-polymerase) with NcoI and XbaI (1x Tango buffer).</p> | + | <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. <a href= "https://2008.igem.org/ wiki/index.php?title=Wiki/Team:Warsaw/ protocols#digest"> Digestion</a> of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).</ p> | + | < 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> |
| | + | <h4>Michał K.</h4> |
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| - | <p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestion</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).</p> | + | <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.4 - 4850 bp, p.5 - 4250 bp).</p>
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| - | <p>7. <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</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. <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.</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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