Protocol for Digestion Prior to droplet digital PCR (ddPCR)

Restriction Enzymes in Droplet Digital PCR

Droplet digital PCR is a method for accurately quantitating copies of DNA or RNA in a sample. Each PCR reaction is separated into thousands or millions of droplets for analysis. Learn more about droplet digital PCR.

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