Team:Warsaw/Calendar-Main/17 June 2008

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<li>Electrophoresis to estimate the concentration of isolated DNA.</li>
<li>Electrophoresis to estimate the concentration of isolated DNA.</li>
<li> Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer). </li>
<li> Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">digest</a> of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer). </li>
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<li> Inoculation of  <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with pACYC177 plasmid into liquid LB + kanamycin. </li>  
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<li> Inoculation of  <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177>pACYC177</a> plasmid into liquid LB + kanamycin. </li>  
</ol></p>
</ol></p>
<h3> Blue/white and rifampicin test </h3>
<h3> Blue/white and rifampicin test </h3>

Revision as of 17:15, 12 October 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. PCR on OmpA_linker and linker_alpha with OmpaL_N and AlphaP_XB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
  2. PCR on OmpA_linker and linker_omega with OmpaL_N and OmegaP_EPB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
  3. Gel electrophoresis of PCR products and gel-out of OmpA_alpha (1000 bp) and OmpA_omega (800 bp).
  4. Electrophoresis to estimate the concentration of isolated DNA.
  5. Overnight digest of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer).
  6. Inoculation of E. coli TOP10 with pACYC177 plasmid into liquid LB + kanamycin.

Blue/white and rifampicin test

Michał L., Ewa, Marcin

Repetition of experiment from 11/06/2008.

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