Team:Warsaw/Calendar-Main/12 May 2008

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(New page: {{WarNotebook}} 1. Restriction digest of pTrc99A-AID with HindIII and NcoI. 2. DNA electrophoresis and isolation of DNA encoding AID from proper band. 3. Restriction digest of pMPM-T5 w...)
 
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<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> construct</h3>
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<h4>Michał K.</h4>
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1. Restriction digest of pTrc99A-AID with HindIII and NcoI.
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<li> <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#electrotransform">electroporation</a> with ligation (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a>).</li>
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2. DNA electrophoresis and isolation of DNA encoding AID from proper band.
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<li>Plating on LB + tetracycline.</li></ol></p>
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3. Restriction digest of pMPM-T5 with HindIII and NcoI, use of alkaline phosphatase (CIAP).
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4. DNA electrophoresis and isolation of pMPM-T5 vector from proper band.
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5. DNA ligation (pMPM-T5+AID vector)
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<html><div align="center"><img src="http://openwetware.org/images/e/e1/UW_AID.jpg" height="340"/></div></html>
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Latest revision as of 16:11, 12 October 2008

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Preparation of pMPMT5+AID construct

Michał K.

  1. E. coli TOP10 electroporation with ligation (pMPMT5+AID).
  2. Plating on LB + tetracycline.

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