AID in transcription fusion works like AID, but induce slower growth than AID;
AID in translation fusion do not induce, or weakly induce rifampicin resistance mutation, it induce slower growth than AID

Optimisation of primers from 10.06

Michał K.

1. PCR on pB30D plasmid using OmpaL_N and OmpaP_link primers (elongation length 45s). Fig. 1 and Fig. 2.
2. PCR on pUC19 plasmid using AlphaL_link and AlphaP_XB primers (elongation length 45s). Fig. 2 and Fig. 3.
3. PCR on pUC19 plasmid using OmegaL_link and OmegaP_EPB primers (elongation length 30s). Fig. 3 and Fig. 4.

Each reaction was carried out with 15 and 20 cycles and in temperature gradient from 55 to 75 degrees.




Gradient PCR products: 1, 14 and 27-DNA ladder;
2 to 8 - gradient PCR for Alpha carried out with 15 cycles;
9 to 16 - gradient PCR for Alpha carried out with 20 cycles;
17 to 23 - gradient PCR for Omega carried out with 15 cycles;
24 to 31 - gradient PCR for Omega carried out with 20 cycles;
32 to 38 - gradient PCR for OmpA carried out with 15 cycles;
39 to 46 - gradient PCR for OmpA carried out with 20 cycles.
Product od each variant of PCR is in 7 lanes: 1^st is the lower border annealing temperature (55 degrees), 6^th is the higher annealing temperature (75 degrees) and 7^th is negative control without template DNA.