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Team:Warsaw/Calendar-Main/17 July 2008
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| - | + | <h3>Cloning of protein Z DNA to OmpA constructs</h3><h4>Michał K.</h4> | |
| - | <h3>Cloning of protein Z DNA to OmpA constructs< | + | |
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| - | <li> | + | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation (with CIAP)</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA_alpha</a> with SacI and NotI (BamHI buffer). </li> |
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<li> Gel eloctrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> (4300 bp band). </li> | <li> Gel eloctrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> (4300 bp band). </li> | ||
<li> | <li> | ||
| - | <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragment and Z DNA fragment isolated on | + | <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragment and Z DNA fragment isolated on <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/14_July_2008">14 July</a>. </li> |
| - | <li> Transformation of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li> | + | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li> |
<li> Transformants plating on LB + kanamycin. </li> | <li> Transformants plating on LB + kanamycin. </li> | ||
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| - | <h3>Cloning of protein | + | |
| - | + | <h3>Cloning of protein Z DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> in place of OmpA</h3><h4>Paweł</h4> | |
| - | + | <p><ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> and Z (in <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pGeneart_Z>Geneart vector</a>) with NdeI and NotI (Orange buffer).</li> | |
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of Z (~200 bp band).</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Gel-out</a> of Z (~200 bp band).</li> | ||
| - | <li>Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of Z into digested pET15b-OmpA-omega.</li> | + | <li>Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of Z into digested <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a>.</li> |
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Cloning of protein Z DNA to OmpA constructsMichał K.
Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpAPaweł
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