Team:Warsaw/Calendar-Main/27 June 2008
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- | <h3>Change of the reporter from | + | <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> |
<h4>Piotr, Weronika</h4> | <h4>Piotr, Weronika</h4> | ||
<ol> | <ol> | ||
- | <li>Isolation of | + | <li>Isolation of <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).</li> |
- | + | <li>Digest of <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) with NotI.</li> | |
- | + | ||
- | <li>Digest of | + | |
<li> | <li> | ||
- | Digest of | + | Digest of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with NotI and <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing>dephosphorylation</a> with CIAP. |
</li> | </li> | ||
<li> | <li> | ||
- | Gel electrophoresis and gel out of proper bands | + | Gel electrophoresis and gel-out of proper bands. |
</li> | </li> | ||
<li> | <li> | ||
- | Ligation of | + | Ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with standard 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). |
</li> | </li> | ||
- | <li>Chemotransformation of E.coli TOP10 with ligation products</li> | + | <li>Chemotransformation of E.coli TOP10 with ligation products.</li> |
- | <li>Plating transformants on LB+Amp30+X-gal+IPTG | + | <li>Plating transformants on LB+Amp30+X-gal+IPTG. |
</li> | </li> | ||
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- | <h3>Preparation of constructs | + | <h3>Preparation of constructs: OmpA_alpha and OmpA_omega</h3><h4>Michał K.</h4> |
<p><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Clean-up</a> of OmpA_alpha and OmpA_omega digest products. </p> | <p><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Clean-up</a> of OmpA_alpha and OmpA_omega digest products. </p> | ||
- | <h3>Cloning | + | <h3>Cloning alpha-A and omega-A fusions on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a></h3> |
<h4>Michał L., Ewa, Marcin</h4> | <h4>Michał L., Ewa, Marcin</h4> | ||
<ol> | <ol> | ||
<li>Gel electophoresis of PCL products (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/27_June_2008#fig1">Fig. 1</a>).</li> | <li>Gel electophoresis of PCL products (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/27_June_2008#fig1">Fig. 1</a>).</li> | ||
- | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of proper bands (alpha-A: | + | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of proper bands (alpha-A: 1100 bp and omega-A: 750 bp).</li> |
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Restriction digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a> vector and the PCL products with NotI and SacI (BamHI buffer) for 2 hours.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Restriction digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pKSII%2B>pKS</a> vector and the PCL products with NotI and SacI (BamHI buffer) for 2 hours.</li> | ||
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> 1 hour.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> 1 hour.</li> | ||
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/ea/PCL_OmegaA_WAW.jpg" width=200/></a> | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/ea/PCL_OmegaA_WAW.jpg" width=200/></a> | ||
- | <var><b>Fig. 1.</b>PCL product - | + | <var><b>Fig. 1.</b>PCL product - Alpha-A fusion.</var> |
</html> | </html> |
Latest revision as of 19:33, 28 October 2008
Change of the reporter from pZC320 with B-galactosidase to GFP or RFPPiotr, Weronika
Michał K.Clean-up of OmpA_alpha and OmpA_omega digest products. Cloning alpha-A and omega-A fusions on pKSMichał L., Ewa, Marcin
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