WarmStart Afu Uracil-DNA Glycosylase (UDG) is inactive at lower temperatures and does not interfere with SDA Amplification below 42°C. In contrast, the Afu Uracil-DNA Glycosylase (UDG) is active <42°C and completely inhibits SDA amplification at these temperatures. Additionally, WarmStart Afu Uracil-DNA Glycosylase (UDG) is fully active at 65°C and can excise incorporated uracils from the SDA product once the temperature is raised to 65°C.
Carryover contamination prevention by WarmStart Afu Uracil-DNA Glycosylase (UDG) requires two parts:
- incorporation of dUTP at lower temperatures (<42°C) by a DNA polymerase during amplicon generation, and
- post-amplification excision of those uracils in amplified products and amplicon destruction catalyzed by a WarmStart Afu Uracil-DNA Glycosylase (UDG) at higher temperatures (~65°C).
