HomeFAQsWhy is there a 60°C, 5 min heat step at the end of the assembly reaction?
FAQ: Why is there a 60°C, 5 min heat step at the end of the assembly reaction?
The final incubation step at 60°C favors BsaI cutting, in the absence of DNA ligation. Digesting any uncut or cut/religated destination plasmid still present in the assembly reactions reduces background.
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