Team:Warsaw/Calendar-Main/20 October 2008

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<h3>Preparation of pT7_alpha_link</h3>
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a></h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<p>Inoculation of colonies from plate with ligation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> (from 30 September) + pT7_alpha to LB with kanamycin.</p>
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<p>Inoculation of colonies from plate with ligation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a> to LB with kanamycin.</p>
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103002>AID under pBAD/araC (BBa_K103002)</a></h3>
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<h4>Michał K.</h4>
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<p> Inoculation of colonies from plate with ligation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/Part:BBa_K103002>AID under pBAD/araC (BBa_K103002)</a> to LB with ampicillin. </p>
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103014>OmpA_linker_alpha_linker (BBa_K103014)</a></h3>
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<h4>Michał K., 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#digest>Digest</a> of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> +
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<a href=http://partsregistry.org/Part:BBa_K103017>OmpA_linker_alpha_linker under Plac (BBa_K103017)</a> 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 <a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a> lane and 3000 bp for <a href=http://partsregistry.org/Part:pSB2K3>PSB2K3 (+ alpha linker)</a> lane.
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<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/20_October_2008#fig1">Fig. 1</a>. </li>
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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>
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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>
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<h3>Preparation of BioBricks</h3>
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<li> Tranformants plating on LB with kanamycin. </li>
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<h4>Michał K.</h4>
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<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
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3kb(????)+pLac_OmpA_alpha and RFP(????)+OmpA  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><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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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/5/58/Go_20_10_2008.jpg"></a>
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<var><b>Fig. 1.</b>EcoRI/SacI digests of plasmids pSB2K3 + OmpA_linker_alpha_linker under Plac (BBa_K103017) and OmpA_linker (BBa_K103006): <br>
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1. Marker<br>
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2.EcoRI/SacI digest of pSB2K3+OmpA_linker (BBa_K103006)<br> 
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3.EcoRI/SacI digest of pSB2K3+OmpA_linker_alpha_linker under Plac (BBa_K103017)<br></var>
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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>
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</ol>
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<li> Tranformants planting on LB with kanamycin. </li>
 
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</ol>
 
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Latest revision as of 16:48, 28 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 AID under pBAD/araC (BBa_K103002)

Michał K.

Inoculation of colonies from plate with ligation of pSB1A3 + AID under pBAD/araC (BBa_K103002) to LB with ampicillin.

Preparation of OmpA_linker_alpha_linker (BBa_K103014)

Michał K., Piotr

  1. Digest of pSB2K3 + OmpA_linker_alpha_linker under Plac (BBa_K103017) 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 PSB2K3 (+ alpha linker) lane. Fig. 1.
  3. Ligation of isolated DNA fragments.
  4. Transformation of TOP10 with above ligation.
  5. Tranformants plating on LB with kanamycin.
    6. Fig. 1.EcoRI/SacI digests of plasmids pSB2K3 + OmpA_linker_alpha_linker under Plac (BBa_K103017) and OmpA_linker (BBa_K103006):
    1. Marker
    2.EcoRI/SacI digest of pSB2K3+OmpA_linker (BBa_K103006)
    3.EcoRI/SacI digest of pSB2K3+OmpA_linker_alpha_linker under Plac (BBa_K103017)

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