Team:Warsaw/Calendar-Main/17 September 2008

From 2008.igem.org

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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlfaPSpe">AlfaPSpe</a>
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlfaPSpe">AlfaPSpe</a>
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primers (elongation length 45s). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_June_2008#fig2">Fig. 2</a> and <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_June_2008#fig3">Fig. 3</a>.</li>
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primers (elongation length 60s). </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=http://www.fermentas.com/techinfo/nucleicacids/mappuc1819.htm>pUC19</a> plasmid using
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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
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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>
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegPSpe">OmegPSpe</a>
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primers (elongation length 30 s). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_June_2008#fig3">Fig. 3</a> and <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_June_2008#fig4">Fig. 4</a>.</li>
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primers (elongation length 60s). </li>
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<p>Each reaction was carried out with 15 and 20 cycles and in temperature gradient from 55 to 75 &deg;C.</p>
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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
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegPSpe">OmegPSpe</a>
 +
primers (elongation length 60s). </li>
 +
<p>Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 &deg;C.</p>
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Revision as of 16:51, 18 October 2008

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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).
  2. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using LinLSXNE and AlfaPSpe primers (elongation length 60s).
  3. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using LinLSXNE and OmegPSpe primers (elongation length 60s).
  4. PCR on pACYC177+OmpA_omega_deltaA_alpha plasmid using LinLSXNE and OmegPSpe primers (elongation length 60s).

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

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