Team:Warsaw/Calendar-Main/30 September 2008

From 2008.igem.org

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<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer). </li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer). </li>
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha with BamHI and BglII (BamHI buffer). </li>
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha with BamHI and BglII (BamHI buffer). </li>
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<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li> </ol>
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<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li>  
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<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">digest</a> of purified pLac_OmpA_omega fragment (from previous day) with EcoRI (EcoRI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li>
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</ol>
<h4>Piotr</h4>
<h4>Piotr</h4>

Revision as of 14:10, 24 October 2008

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Mutagenesis of protein A

Paweł

Results of sequencing: unfortunately all sequences where wild-type.

Preparation of BioBricks

Michał K.

  1. Isolation of HQs(??????) plasmids.
  2. Digest of isolated plasmids with EcoRI and BcuI (BamHI buffer).
  3. Digest of pACYC177+OmpA_omega_deltaA_alpha with BamHI and BglII (BamHI buffer).
  4. Gel elctrophoresis and gel-out of proper band: ??????.
  5. Overnight digest of purified pLac_OmpA_omega fragment (from previous day) with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment (3 hr).

Piotr

  1. E. coli TOP10 electroporation with overnight ligations (RFP(??????) + Z and RFP(????) + OmpA_link).
  2. Plating on LB + ampicillin.