Protocol for Digestion Prior to droplet digital PCR (ddPCR)
- Setup restriction enzyme digests in the recommended NEBuffer
- Use 10 units of restriction enzyme per microgram of DNA sample
- Incubate at incubation temperature for 5–60 minutes as recommended for each enzyme
- Enzyme can be heat inactivated if desired, but this is not required
- No cleanup is necessary after digestion; sample can be directly added to ddPCR reactions
- Avoid carrying over more than a 1/10 amount of the restriction digest mixture to the ddPCR reaction