Team:Warsaw/Calendar-Main/30 September 2008

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Revision as of 18:02, 24 October 2008


	Attribution analyzer / Interactive list of topics

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

Isolation of HQs(??????) plasmids.
Digest of isolated plasmids with EcoRI and BcuI (BamHI buffer).
Digest of pACYC177+OmpA_omega_deltaA_alpha with BamHI and PstI (BamHI buffer).
Gel elctrophoresis and gel-out of proper band: ??????.
Digest of link_omega fragment (PCR from previous day) with EcoRI and BcuI (BamHI buffer)
Digest of OmpA_omega with BglII and PstI (Orange buffer).
Clean-up of above digest reactions.
Overnight digest of purified pLac_OmpA_omega fragment (from previous day) with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment (3 hr).

E. coli TOP10 transformation with overnight ligations (RFP(??????) + Z and RFP(????) + OmpA_link).
Plating on LB + ampicillin.
Inoculation of bacterias received from iGEM HQs ( ).

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-<h3>Mutagenesis of protein A</h3><h4>Paweł</h4>
+<h3>Mutagenesis of protein A</h3>
+<h4>Paweł</h4>
 <p>Results of sequencing: unfortunately all sequences where wild-type.</p>
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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>
 </ol>
+<h3></h3>
 <h4>Piotr</h4>
 <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/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformation</a> with overnight ligations (RFP(??????) + Z and RFP(????) + OmpA_link). </li>