Team:Warsaw/Calendar-Main/17 October 2008

Preparation of OmpA-linker-omega-linker (BBa_K103016) Michał K. Isolation of plasmids from cultures inoculated on previous day pACYC177 + OmpA-linker-omega-linker (BBa_K103016).

Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found Fig. 1.</li></ol>

<img src="http://2008.igem.org/wiki/images/7/75/Traw_17_10_2008.jpg"></a> Fig. 1.EcoRI/PstI digests of plasmids pACYC177+OmpA-linker-omega-linker (BBa_K103016). 1. Marker 2-9. EcoRI/PstI digests of pACYC177+OmpA-linker-omega-linker (BBa_K103016)

Preparation of alpha_linker under PT7 (BBa_K103019)</a> Michał K., Piotr <ol>

Gel electrophoresis of over night digest (<A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying alpha_linker under PT7 (BBa_K103019)</a>) and gel-out</a> of proper band - 800 bp. Fig. 2</a>. </li> Ligation</a> of isolated DNA fragments: pSB2K3</a> (from 30 September</a>) + alpha_linker under PT7 (BBa_K103019)</a>.</li>  Transformation</a> of TOP10</a> with above ligation.</li>  Plating on LB with kanamycin.</li></ol>

<img src="http://2008.igem.org/wiki/images/e/e5/Go_17_10_2008.jpg"></a> Fig. 2.EcoRI/BcuRI digests of BBa_K103019. 1. Marker 2. EcoRI/BcuI digest of BBa_K103019

Preparation of AID under pBAD/araC (BBa_K103002)</a> Piotr <ol> <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> <li> Plating on LB with ampicillin.</li>

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