Team:Warsaw/Calendar-Main/1 July 2008

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<h3>Preparation of constructs with OmpA protein fusions</h3>
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<li> PCR on pBD (jaki numerek?) plasmid with
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<p>'''Preparation of constructs with OmpA protein fusions'''<br>
 
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1. PCR on pBD (jaki numerek?) plasmid with
 
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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>
<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, annealing length 45s, annealing temperature 63 degrees)<br>
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  primers (15 cycles, annealing duration 45 s, annealing temperature 63&deg;C) </li>
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2. PCR on pUC19 plasmid with  
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<li> PCR 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>
<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, annealing length 45s, annealing temperature 63 degrees)<br>
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  primers (20 cycles, annealing length 45s, annealing temperature 63&deg;C)</li>
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3. PCR on pUC19 plasmid with  
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<li> PCR on pUC19 plasmid with  
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<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>
<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, annealing duration 30 s, annealing temperature 58&deg;C)</li>
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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. <br>
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<li> Gel electrphoresis 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</li>
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</ol></p>
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primers (20 cycles, annealing length 30s, annealing temperature 58 degrees)<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. <br>
 
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4. Gel electrphoresis of PCR products and gel-out of proper bands</p>
 
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Revision as of 16:11, 1 October 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 duration 45 s, annealing temperature 63°C)
  2. PCR on pUC19 plasmid with AlphaL_link and AlphaP_XB primers (20 cycles, annealing length 45s, annealing temperature 63°C)
  3. PCR on pUC19 plasmid with OmegaL_link and OmegaP_EPB primers (20 cycles, annealing duration 30 s, annealing temperature 58°C)
    4. 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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