Team:Warsaw/Calendar-Main/1 July 2008

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<ol>
<ol>
<li> PCR on pBD (jaki numerek?) plasmid with  
<li> 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 duration 45 s, annealing temperature 63&deg;C) </li>
  primers (15 cycles, annealing duration 45 s, annealing temperature 63&deg;C) </li>
<li> PCR on pUC19 plasmid with  
<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>
-
 
+
  primers (20 cycles, annealing duration 45 s, annealing temperature 63&deg;C)</li>
-
  primers (20 cycles, annealing length 45s, annealing temperature 63&deg;C)</li>
+
<li> PCR on pUC19 plasmid with  
<li> PCR 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>
<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)<br>
-
  primers (20 cycles, annealing duration 30 s, annealing temperature 58&deg;C)</li>
+
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega. </li>
-
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega. <br>
+
<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>
<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>
</ol></p>
</ol></p>

Revision as of 16:12, 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 duration 45 s, 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)
    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