Team:Warsaw/Calendar-Main/17 September 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. 55 °C deltaA
3. 60 °C deltaA
4. 65 °C deltaA
5. 70 °C deltaA
6. 55 °C link_omega
7. 60 °C link_omega
8. 65 °C link_omega
9. 70 °C link_omega
10. 55 °C link_alpha
11. 60 °C link_alpha
12. 65 °C link_alpha
13. 70 °C link_alpha
14. 55 °C OmpA_omega
15. 60 °C OmpA_omega
16. 65 °C OmpA_omega
17. 70 °C OmpA_omega