Team:Warsaw/Calendar-Main/17 September 2008

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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on pACYC177+OmpA_omega_deltaA_alpha plasmid using
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on pACYC177+OmpA_omega_deltaA_alpha plasmid using
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmLNXNEB">OmLNXNEB</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinPBSNP">LinPBSNP</a>
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpLNXNE">OmpLNXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinPBSNP">LinPBSNP</a>
primers (elongation length 90s) to obtain Omp_omega fragment. </li>
primers (elongation length 90s) to obtain Omp_omega fragment. </li>
<p>Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 &deg;C.</p>
<p>Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 &deg;C.</p>

Revision as of 19:10, 21 October 2008

Gallery Bricks Notebook Team Project Home


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Competition tests

Piotr

  1. Plasmid isolation from previous day's cultures.
  2. Digest with SacI and BamHI.
  3. Electrophoresis.

[photo of the gel is to be placed here]

Conclusion: cells with interracting protein survive competition!

MutD5 testing

Emilia

Inoculation of MutD5 into medium with tetracycline.

Optimisation of primers for preparation of parts

Michał K.

  1. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using AL_BNXNE and APSacSpe primers (elongation length 45s) to obtain deltaA fragment.
  2. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using LinLSXNE and AlfaPSpe primers (elongation length 60s) to obtain link_alpha fragment.
  3. PCR on pACYC177 + OmpA_omega plasmid using LinLSXNE and OmegPSpe primers (elongation length 60s) to obtain link_omega fragment.
  4. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using OmpLNXNE and LinPBSNP primers (elongation length 90s) to obtain Omp_omega fragment.

    5. Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 °C.

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