Team:Warsaw/Calendar-Main/20 October 2008

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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> +  
<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> +  
<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>
<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  
+
<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.
<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/20_October_2008#fig1">Fig. 1</a>. </li>
<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/20_October_2008#fig1">Fig. 1</a>. </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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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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