Team:Warsaw/Calendar-Main/23 May 2008

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(New page: {{WarNotebook}} <!-- do not edit above me! --> 1. Digest of pMPM-T5 with NcoI and XbaI 2. Digest of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI 3. Cl...)
 
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1. Digest of pMPM-T5 with NcoI and XbaI
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<h3>Rifampicin test #2</h3>
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<h4>Piotr</h4>
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2. Digest of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI
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<p> Plating cultures from the previous day on rifampicin.</p>
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3. Clean-up of digested products from p.1 and p.2
 
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4. Ligation of digested products from p.1 and p.2
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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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5. Transformation of E.coli TOP10 with ligation product
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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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6. Digest of pMPM-T5+AID with HindIII and SalI
 
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7. Digest of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI
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8. Clean-up of digested products from p.6 and p.7
 
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9. Ligation of digested products from p.6 and p.7
 
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10. Transformation of E.coli TOP10 with ligation product
 
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11. Transformants plating on LB + tetracycline
 
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<img src="http://openwetware.org/images/thumb/6/61/UW_A%2BT.jpg/350px-UW_A%2BT.jpg"/> <img src="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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