Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA
Paweł
- Isolation of plasmids from cultures inocluated on previous day.
- Control digest of isolated plasmids with NdeI and NotI (Orange buffer). Zero positive results obtained - just empty vectors.
- Another overnight ligation of pET15b-OmpA-omega (NdeI/NotI cut) with protein Z DNA.
Cloning omega-A fusion on pKS (second attempt)
Michał L., Ewa, Marcin
We have obtained white colonies of transformants and inoculated 10 of them on liquid LB+Amp100.
Cloning of protein A DNA to OmpA constructsMichał K.
- Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
AL+SacI and AP+NotI.
- Gel electrophoresis (Fig. 1.).
- Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin.
Fig. 1. Colony PCR on transformants with pACYC177_OmpA_alpha_A
Upper and lower:
1. Marker
2-13. colony PCR (pACYC177_OmpA_alpha_A)
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