Team:Warsaw/Calendar-Main/14 October 2008

Preparation of linker_alpha (BBa_K103009) Piotr   Transformation of TOP10 with overnight ligation - pSB2K3 + linker_alpha (BBa_K103009).

 Tranformants plating on LB with kanamycin. 



Preparation of <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a> Michał K. <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day - <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a>.</li> Control <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found. <a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/14_October_2008#fig1">Fig. 1</a>.</li></ol>

<img src="http://2008.igem.org/wiki/images/3/37/Traw_18_10_2008.jpg"></a> Fig. 1.EcoRI/PstI digests of isolated plasmids. 1. Marker 2-8.EcoRI/PstI digests of pACYC177+OmpA-linker-omega-linker plasmids

Preparation of <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a> Michał K. <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day - <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>.

Preparation of <a href=http://partsregistry.org/Part:BBa_K103020>omega_linker under PT7 (BBa_K103020)</a> Michał K. <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103020>omega_linker under PT7 (BBa_K103020)</a>).</li> Control <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. <a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/14_October_2008#fig2">Fig. 2</a>.</li></ol>

<img src="http://2008.igem.org/wiki/images/e/ea/Traw_14_10_2008.jpg" width=200></a> Fig. 2.EcoRI/PstI digests of isolated pSB2K3+omega_linker under PT7 (BBa_K103020) plasmids 1. Marker 2-4. Digested plasmids pSB2K3+omega_linker under PT7 (BBa_K103020)

Preparation of <a href=http://partsregistry.org/Part:BBa_K103002>AID under pBAD/araC (BBa_K103002)</a> Michał K.  <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="http://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with overnight ligation - <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>.</li>

 Tranformants plating on LB with ampicillin. </li>

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Preparation of <a href=http://partsregistry.org/Part:BBa_K103017>OmpA_linker_alpha_linker under Plac (BBa_K103017)</a> Piotr 

 <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="http://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with overnight ligation - <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> (with OmpA_linker and lactose promoter) + alpha2.</li>

 Tranformants plating on LB with kanamycin. </li>

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