Team:Warsaw/Calendar-Main/20 June 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. Isolation of plasmids from cultures inoculated on previous day.
  2. Control digest of isolated plasmids with PstI (Orange buffer).
  3. Gel electrophoresis (Fig. 1 and 2, no proper colonies founded).

Preparation of alpa-A and omega-A fusions

Michał L., Ewa, Marcin

PCR
ProductTemplatePrimersProduct length
linker-ApDRIVE-TapTagAL+link10+homo2 and AP+NotI 470 bp
alpha-linkerpUC19 AlphaL+SacI and AlphaP+link10+homo2600 bp
omega-linkerpUC19OmegaL+SacI and OmegaP+link10+homo2400 bp



Universal PCR program for A, alpha and omega
TemperatureTime
94°C4:00
94°C0:3028 cycles
50°C0:45
72°C2:00
72°C10:00
4°Cinfinite

Fig. 1. Lanes 1 to 8 - control digest of 8 clones carrying probably OmpA_alpha fusion with PstI. Fig. 2. Lanes 1 to 8 - control digest of 8 clones carrying probably OmpA_omega fusion with PstI.





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