Team:Warsaw/Calendar-Main/21 October 2008

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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day (<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>).</li>
<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day (<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>).</li>
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<li>Control <a href=https://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.</li></ol>
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<li>Control <a href=https://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="https://2008.igem.org/Team:Warsaw/Calendar-Main/21_October_2008#fig1">Fig. 1</a>.</li></ol>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/1/11/Traw2_19_10_2008.jpg" width=200></a>
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<var><b>Fig. 1.</b> EcoRI/PSTI digest of pT7_alpha_link </var>
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<h3>Preparation of AraC+pBAD+AID</h3>
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1-3. EcoRI/PstI digests of pT7_alpha_link<br> 4. Marker<br></var>
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103002>AID under pBAD/araC (BBa_K103002)</a></h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<ol><li> Inoculation of few more colonies from plate with ligation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + AraC+pBAD+AID to LB with ampicillin. </li>
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<ol><li> Inoculation of few more 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. </li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day - <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + AraC+pBAD+AID.</li>
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<li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures 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>
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<li>Control <a href=https://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.</li>
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<li>Control <a href=https://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="https://2008.igem.org/Team:Warsaw/Calendar-Main/21_October_2008#fig2">Fig. 2</a>.</li>
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</ol>
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/a/ac/Traw4_16_10_2008.jpg"></a>
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<var><b>Fig. 2.</b> EcoRI/PstI digests of BBa_K103002  <br>
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1. Marker <br>
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2-6. EcoRI/PstI digests of BBa_K103002 <br></var>
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<h3>Preparation of OmpA-linker-alpha-linker</h3>
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<h3>Preparation of <a href=http://partsregistry.org/Part:BBa_K103014>OmpA_linker_alpha_linker (BBa_K103014)</a></h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<p> Inoculation of colonies from plate with ligation (OmpA_alpha_) to LB with kanamycin. </p>
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<p> Inoculation of colonies from plate with ligation (<a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=<a href=http://partsregistry.org/Part:BBa_K103014>OmpA_linker_alpha_linker (BBa_K103014)</a>) to LB with kanamycin. </p>

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Preparation of OmpA-linker-omega-linker (BBa_K103016)

Piotr

Inoculation of few more colonies from plate with ligation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016) (from 16 October) to LB with kanamycin.

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

  1. Isolation of plasmids from cultures inoculated on previous day (pSB2K3 + alpha_linker under PT7 (BBa_K103019)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. Fig. 1.
Fig. 1. EcoRI/PSTI digest of pT7_alpha_link 1-3. EcoRI/PstI digests of pT7_alpha_link
4. Marker

Preparation of AID under pBAD/araC (BBa_K103002)

Michał K.

  1. Inoculation of few more colonies from plate with ligation of pSB1A3 + AID under pBAD/araC (BBa_K103002) to LB with ampicillin.
  2. Isolation of plasmids from cultures inoculated on previous day - pSB1A3 +AID under pBAD/araC (BBa_K103002).
  3. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found. Fig. 2.
Fig. 2. EcoRI/PstI digests of BBa_K103002
1. Marker
2-6. EcoRI/PstI digests of BBa_K103002

Preparation of OmpA_linker_alpha_linker (BBa_K103014)

Michał K.

Inoculation of colonies from plate with ligation (pSB2K3 + OmpA_linker_alpha_linker (BBa_K103014)) to LB with kanamycin.

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