Home FAQs Why do assembly reactions end with a 5 minute, 60°C incubation step?

FAQ: Why do assembly reactions end with a 5 minute, 60°C incubation step?

The final incubation step at 60°C favors Type IIS restriction enzyme cutting, in the absence of DNA ligation. Digesting any uncut or cut/religated destination plasmid still present in the assembly reactions reduces background.