Team:Warsaw/Calendar-Main/25 September 2008

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<h3>Preparation of BioBricks</h3>
<h3>Preparation of BioBricks</h3>
<h4>Piotr</h4>
<h4>Piotr</h4>
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<ol><li> <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#electrotransform">electroporation</a> with overnight ligation of RFP(??????) and deltaA DNA fragments. </li>
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<ol><li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <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 on colonies from plates with transformations OmpA_alpha (45 s of elongation step). </li>
<li>Plating on LB + ampicillin.</li>
<li>Plating on LB + ampicillin.</li>

Revision as of 20:07, 22 October 2008

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Mutagenesis of protein A

Paweł

Treatment of mutageneses as on 23rd September.

Preparation of alpha_A construct

Antoni

  1. PCR on alpha_linker and linker_A with AlphaL+SacI and AP+NotI primers and 10% DMSO (30 cycles, elongation 60 s, annealing temperature 72°C).
  2. Gel electrophoresis of PCR products and gel-out of proper band (1019 bp).
  3. Measurment of concentration of both isolated products.

Preparation of BioBricks

Piotr

  1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations OmpA_alpha (45 s of elongation step).
  2. Plating on LB + ampicillin.

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