Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
- Digestion is recommended whenever DNA input is greater than 75 ng
- Setup reactions at room temperature to allow digestion
- 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
- After setup, simply continue droplet generation as normal.
- Restriction enzyme will be inactivated during first PCR denaturation step