Team:Warsaw/Calendar-Main/30 June 2008

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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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<h4>Piotr, Weronika</h4>
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<li>There were blue and white colonies but no red or green in UV.</li>
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<li>Inoculation of liquid LB medium + Amp with white colonies.</li>
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<li>Inoculation of 10ml LB with E.coli TOP10 carrying <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a>.</li>
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<li>Inoculation of E.coli TOP10 carrying <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A2>pSB1A2</a> standard plasmids carrying parts: <A href=http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840>BBa_E0840</a>(GFP genetrator) and <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450>BBa_J04450</a> (RFP generator).
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<p>'''Optimisation of primers from 19.06'''<br>
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1. PCR on pBD (jaki numerek?) plasmid with
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>
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primers (annealing length 45s)<br>
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2. PCR on pUC19 plasmid with
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaL_link">AlphaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaP_XB">AlphaP_XB</a>
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primers (annealing length 45s)<br>
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3. PCR on pUC19 plasmid with
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaL_link">OmegaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaP_EPB">OmegaP_EPB</a>
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primers (annealing length 30s)<br>
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Each reaction was carried out with 15 and 20 cycles and in temperature gradient from 55 to 75 degrees</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>Inoculation of liquid LB medium with 6 transformant colonies.</p>
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</ol>
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<h3>Preparation of constructs on pET15b: OmpA_alpha and OmpA_omega</h3><h4>Paweł</h4>
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<p> Inoculation of <i> E. coli</i> with <A href=http://www.emdbiosciences.com/docs/docs/PROT/TB045.pdf>pET15b</a> plasmid on LB + ampicillin.</p>
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Latest revision as of 14:03, 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 in UV.
  2. Inoculation of liquid LB medium + Amp with white colonies.
  3. Inoculation of 10ml LB with E.coli TOP10 carrying pZC320.
  4. Inoculation of E.coli TOP10 carrying pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator).

Cloning alpha-A and omega-A fusions on pKS

Michał L., Ewa, Marcin

Inoculation of liquid LB medium with 6 transformant colonies.

Preparation of constructs on pET15b: OmpA_alpha and OmpA_omega

Paweł

Inoculation of E. coli with pET15b plasmid on LB + ampicillin.

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