From 2008.igem.org
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| - | <h3>Preparation of constructs with OmpA protein fusions<br> | + | |
| - | Michał K.</h3>
| + | <h3>Preparation of constructs with OmpA protein fusions<br>Paweł</h3> |
| - | <p> | + | <p><ol><li> Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/ igem_notebook. htm# digest"> digest</a> of pET15b plasmid with NdeI and BamHI, <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols# removing"> dephosphorylation with CIAP</a>.</li></ol></p> |
| - | <ol> | + | |
| - | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on pB30D plasmid 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 (15 cycles, elongation duration 45 s, annealing temperature 63°C). </li>
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| - | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on pUC19 plasmid with
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| - | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaL_link">AlphaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaP_XB">AlphaP_XB</a>
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| - | primers (20 cycles, elongation duration 45 s, annealing temperature 63°C).</li>
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| - | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols# pcr"> PCR</a> on pUC19 plasmid with | + | |
| - | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaL_link">OmegaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaP_EPB">OmegaP_EPB</a>
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| - | primers (20 cycles, elongation duration 30 s, annealing temperature 58°C).<br>
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| - | As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega. </li>
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| - | <li> Gel electrophoresis of PCR products and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands (OmpA_linker - 500 bp, linker_alpha - 600 bp and linker_omega - 350 bp).</li>
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| - | </ol></p> | + | |
| | </html> | | </html> |
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Revision as of 21:25, 9 October 2008
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