Team:Warsaw/Calendar-Main/1 July 2008

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<p>'''Preparation of constructs with OmpA protein fusions'''<br>
<p>'''Preparation of constructs with OmpA protein fusions'''<br>
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1. PCR on pBD (jaki numerek?) plasmid with ompaL_N and ompaP_link primers (15 cycles, annealing length 45s, annealing temperature 63 degrees)<br>
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1. PCR on pBD (jaki numerek?) plasmid with  
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2. PCR on pUC19 plasmid with alphaL_link and alphaP_XB primers (20 cycles, annealing length 45s, annealing temperature 63 degrees)<br>
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<html>
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3. PCR on pUC19 plasmid with omegaL_link and omegaP_EPB primers (20 cycles, annealing length 30s, annealing temperature 58 degrees)<br>
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<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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</html>
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primers (15 cycles, annealing length 45s, annealing temperature 63 degrees)<br>
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2. PCR on pUC19 plasmid with  
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<html>
 +
<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>
 +
</html>
 +
primers (20 cycles, annealing length 45s, annealing temperature 63 degrees)<br>
 +
3. PCR on pUC19 plasmid with  
 +
<html>
 +
<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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</html>
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primers (20 cycles, annealing length 30s, annealing temperature 58 degrees)<br>
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega. <br>
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega. <br>
4. Gel electrphoresis of PCR products and gel-out of proper bands</p>
4. Gel electrphoresis of PCR products and gel-out of proper bands</p>

Revision as of 14:28, 26 September 2008

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Preparation of constructs with OmpA protein fusions
1. PCR on pBD (jaki numerek?) plasmid with OmpaL_N and OmpaP_link primers (15 cycles, annealing length 45s, annealing temperature 63 degrees)
2. PCR on pUC19 plasmid with AlphaL_link and AlphaP_XB primers (20 cycles, annealing length 45s, annealing temperature 63 degrees)
3. PCR on pUC19 plasmid with OmegaL_link and OmegaP_EPB primers (20 cycles, annealing length 30s, annealing temperature 58 degrees)
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega.
4. Gel electrphoresis of PCR products and gel-out of proper bands

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