Team:Warsaw/Calendar-Main/17 September 2008

From 2008.igem.org

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<p><ol><li>Plasmid isolation from previous day's cultures.</li><li>Digest with SacI and BamHI.</li><li>Electrophoresis.</li></ol></p>
<p><ol><li>Plasmid isolation from previous day's cultures.</li><li>Digest with SacI and BamHI.</li><li>Electrophoresis.</li></ol></p>
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[photo of the gel is to be placed here]
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[photo of gel goes here!]
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<p>Conclusion: cells with interracting protein survive competition!</p>
<p>Conclusion: cells with interracting protein survive competition!</p>
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primers (elongation length 90s) to obtain OmpA_omega fragment. </li>
primers (elongation length 90s) to obtain OmpA_omega fragment. </li>
<p>Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 &deg;C (four reactions).</p>
<p>Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 &deg;C (four reactions).</p>
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<li> Gel electrophoresis of PCR products.</li>
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<li> Gel electrophoresis of PCR products <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_September_2008#fig1">Fig. 1</a>.</li>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/9/9e/Grad_17_09.jpg" width=300/></a><var><b>Fig. 1. Gradient PCR (temperatures:55-75 &deg;C)</b><br>
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1. Marker<br>
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2. 60 &deg;C deltaA<br>
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3. 65 &deg;C deltaA<br>
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4. 70 &deg;C deltaA<br>
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5. 75 &deg;C deltaA<br>
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6. 60 &deg;C omega<br>
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7. 65 &deg;C omega<br>
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8. 70 &deg;C omega<br>
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9. 75 &deg;C omega<br>
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10. 60 &deg;C alpha<br>
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11. 65 &deg;C alpha<br>
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12. 70 &deg;C alpha<br>
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13. 60 &deg;C alpha<br>
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14. 65 &deg;C OmpA_omega<br>
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15. 70 &deg;C OmpA_omega<br>
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16. 75 &deg;C OmpA_omega<br>
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17. 75 &deg;C OmpA_omega<br></var>
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Revision as of 22:01, 26 October 2008

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'Hunter and prey' system tests: Competition tests

Piotr

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

[photo of gel goes 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_ΔA_alpha plasmid using AL_BNXNE and APSacSpe primers (elongation length 45s) to obtain ΔA fragment.
  2. PCR on pACYC177+OmpA_omega_ΔA_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_ΔA_alpha plasmid using OmpLNXNE and LinP_BS primers (elongation length 90s) to obtain OmpA_omega fragment.
  5. Each reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 °C (four reactions).

  6. Gel electrophoresis of PCR products Fig. 1.
Fig. 1. Gradient PCR (temperatures:55-75 °C)
1. Marker
2. 60 °C deltaA
3. 65 °C deltaA
4. 70 °C deltaA
5. 75 °C deltaA
6. 60 °C omega
7. 65 °C omega
8. 70 °C omega
9. 75 °C omega
10. 60 °C alpha
11. 65 °C alpha
12. 70 °C alpha
13. 60 °C alpha
14. 65 °C OmpA_omega
15. 70 °C OmpA_omega
16. 75 °C OmpA_omega
17. 75 °C OmpA_omega