Team:Warsaw/Calendar-Main/20 October 2008

From 2008.igem.org

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<h3>Preparation of BioBricks</h3>
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<h3>Preparation of OmpA_alpha_</h3>
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<h4>Michał K.</h4>
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<h4>Michał K., Piotr</h4>
<ol>
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<li> Inoculation of colonies from plate with ligation of 5kb(?????) + pT7_alpha to LB with kanamycin and RFP(?????) + AraC+pBAD+AID to LB with ampicillin. </li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of  
<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of  
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3kb(????)+pLac_OmpA_alpha and RFP(????)+OmpA  with EcoRI and SacI (BamHI buffer). </li>
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pLac_OmpA_alpha and <a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a> with EcoRI and SacI (BamHI buffer). </li>
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<li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band 500 bp for RFP(????)+OmpA and 3000 bp for 3kb(????)+pLac_OmpA_alpha. </li>
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<li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band 500 bp for <a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a> lane and 3000 bp for pLac_OmpA_alpha lane. </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of DNA fragments: 3kb(?????)_ alpha + Omp.</li></ol>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments.</li></ol>
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<h3></h3>
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<h4>Piotr</h4>
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<ol>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li>
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Revision as of 12:30, 27 October 2008

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Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

Inoculation of colonies from plate with ligation of pSB2K3 + alpha_linker under PT7 (BBa_K103019) to LB with kanamycin.

Preparation of AraC+pBAD+AID

Michał K.

Inoculation of colonies from plate with ligation of pSB1A3 + AraC+pBAD+AID to LB with ampicillin.

Preparation of OmpA_alpha_

Michał K., Piotr

  1. Digest of pLac_OmpA_alpha and OmpA-linker (BBa_K103006) with EcoRI and SacI (BamHI buffer).
  2. Gel electrophoresis and gel-out of proper band 500 bp for OmpA-linker (BBa_K103006) lane and 3000 bp for pLac_OmpA_alpha lane.
  3. Ligation of isolated DNA fragments.
  • Transformation of TOP10 with above ligation.
  • Tranformants planting on LB with kanamycin.

    • Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/20_October_2008"