Team:Warsaw/Calendar-Main/16 October 2008

Preparation of linker_alpha (BBa_K103009) Michał K.  Isolation of plasmids from culture inoculated on previous day. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. Fig. 1.

 Fig. 1.EcoRI/PstI digests of isolated plasmids 1. Marker 2-7. Digests of pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) 8-11. Digests of pSB2K3 carrying alpha_linker under pLac 11-17. Digests of pSB2K3 carrying linker_alpha

Preparation of OmpA-linker-omega-linker (BBa_K103016)</a> Michał K. Inoculation of colonies from plate with new ligation of <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> to LB with kanamycin.

Preparation of <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a> Michał K. Control <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid - <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> fragment (<a href=http://2008.igem.org/Team:Warsaw/Calendar-Main/14_October_2008>14 October</a>) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. <a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/16_October_2008#fig1">Fig. 1</a>.</li> Overnight <a href="http://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> <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> with EcoRI and BcuI (BamHI buffer). </li> </ol>

<img src="http://2008.igem.org/wiki/images/f/f7/Traw_16_10_2008.jpg"> Fig. 1.EcoRI/PstI digests of isolated plasmids 1. Marker 2-7. Digests of pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) 8-11. Digests of pSB2K3 carrying alpha_linker under pLac 11-17. Digests of pSB2K3 carrying linker_alpha

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#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<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> 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/16_October_2008#fig2">Fig. 2</a>.</li>  Repetition of overnight <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <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></ol>

<img src="http://2008.igem.org/wiki/images/3/3c/Traw2_15_10_2008.jpg"></a> Fig. 2.EcoRI/PstI digests of isolated plasmids (pSB1A3+AID under pBAD/araC (BBa_K103002) 1. Marker 2-9. EcoRI/PstI digests of isolated plasmids

Preparation of <a href=http://partsregistry.org/Part:BBa_K103017>OmpA_linker_alpha_linker under Plac (BBa_K103017)</a> Michał K.  <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day - <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></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 - we found good clones. <a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/16_October_2008#fig1">Fig. 1</a>.</li>

</ol>

<img src="http://2008.igem.org/wiki/images/f/f7/Traw_16_10_2008.jpg" width=300></a> Fig. 1.EcoRI/PstI digests of isolated plasmids 1. Marker 2-7. Digests of pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) 8-11. Digests of pSB2K3 carrying alpha_linker under pLac 11-17. Digests of pSB2K3 carrying linker_alpha