Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)

  1. Digestion is recommended whenever DNA input is greater than 75 ng 
  2. Setup reactions at room temperature to allow digestion 
  3. Prepare reaction mixes as you would for a standard ddPCR reaction. Add 0.5–1 μL of each restriction enzyme (5–20 units, depending on enzyme concentration) to the reaction mixture 
  4. After setup, simply continue droplet generation as normal. 
  5. Restriction enzyme will be inactivated during first PCR denaturation step