Team:Warsaw/Calendar-Main/9 May 2008

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<li> Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with HindIII and NcoI (1x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</li>
<li> Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with HindIII and NcoI (1x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</li>
<li> DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> vector from proper band (4250 bp).</li>
<li> DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> vector from proper band (4250 bp).</li>
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<img src="https://static.igem.org/mediawiki/2008/0/0f/Gel_out_pmpm_WAW.jpg" width=300/>
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<var>1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII</var>
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<li>Electrophoresis to estimate the concentration of isolated DNA.</li>
<li>Electrophoresis to estimate the concentration of isolated DNA.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments. </li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments. </li>
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<img src="https://static.igem.org/mediawiki/2008/8/89/Gel_out_aid_WAW.jpg" width=300/>
<img src="https://static.igem.org/mediawiki/2008/8/89/Gel_out_aid_WAW.jpg" width=300/>
<var>1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII</var>
<var>1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII</var>
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<img src="https://static.igem.org/mediawiki/2008/0/0f/Gel_out_pmpm_WAW.jpg" width=300/>
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<var>1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII</var>
<div align="center"><img src="http://openwetware.org/images/e/e1/UW_AID.jpg" height="340"/></div>
<div align="center"><img src="http://openwetware.org/images/e/e1/UW_AID.jpg" height="340"/></div>
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Revision as of 08:49, 11 October 2008

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Preparation of pMPMT5+AID construct

Michał K.

  1. Restriction digest of pTrc99A-AID with HindIII and NcoI (1x Tango buffer).
  2. DNA electrophoresis and isolation of DNA encoding AID from proper band (600 bp).
  3. Restriction digest of pMPM-T5 with HindIII and NcoI (1x Tango buffer), dephosphorylation (CIAP).
  4. DNA electrophoresis and isolation of pMPM-T5 vector from proper band (4250 bp).
  5. Electrophoresis to estimate the concentration of isolated DNA.
  6. Ligation of isolated DNA fragments.

1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII






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