Team:Warsaw/Calendar-Main/3 July 2008

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<h3>Preparation of constructs with OmpA protein fusions
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<h3>Change of the reporter from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with B-galactosidase to GFP or RFP</h3>
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<br>Paweł</h3>
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<h4>Piotr, Weronika</h4>
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<li>There were blue and white colonies but no red or green fluorescence in UV.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of overnight digest.</li>
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<li>Inoculation of liquid LB medium + Amp with white colonies.</li>
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<li> <A HREF="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</A> of OmpA_alfa and pET15b. </li>
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<li> <A HREF="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</A> of OmpA_omega and pET15b.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> TOP10 strain with ligation products. </li>
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<li> Transformants plating on LB + ampicillin.</li></ol>
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<h3>Preparation of constructs on pET15b: OmpA_alpha and OmpA_omega</h3>
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<h4>Paweł</h4>
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Colonies from each transformant strain from previous day (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+OmpA_alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a>) were inoculated to liquid LB + ampicillin.</p>
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<h3>Cloning alpha-A and omega-A fusions on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a></h3>
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<h4>Michał L., Ewa, Marcin</h4>
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<p>We have received sequencing results: A is OK but in case of Alpha-A and Omega-A the result is weird. It seems that there were error in synthesis of OmegaP+link10+homo2 and AlphaP+link10+homo2 primers which causes frameshift and cloning of these fusions has to be repeated from the beginning.</p>
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Latest revision as of 14:07, 28 October 2008

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Change of the reporter from pZC320 with B-galactosidase to GFP or RFP

Piotr, Weronika

  1. There were blue and white colonies but no red or green fluorescence in UV.
  2. Inoculation of liquid LB medium + Amp with white colonies.

Preparation of constructs on pET15b: OmpA_alpha and OmpA_omega

Paweł

Colonies from each transformant strain from previous day (pET15b+OmpA_alpha and pET15b+OmpA_omega) were inoculated to liquid LB + ampicillin.

Cloning alpha-A and omega-A fusions on pKS

Michał L., Ewa, Marcin

We have received sequencing results: A is OK but in case of Alpha-A and Omega-A the result is weird. It seems that there were error in synthesis of OmegaP+link10+homo2 and AlphaP+link10+homo2 primers which causes frameshift and cloning of these fusions has to be repeated from the beginning.

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