Restriction Digest
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Protocol
- Check the NEB (or other supplier) website to confirm enzyme compatibility
- In a PCR tube add the reaction components according to the table listed below
- Mix reaction gently and spin down briefly with benchtop centrifuge
- Incubate at 37°C for 30 min if time-saver qualified, 60 minutes if not
Example Reaction
Component | Volume |
ddH2O | Up to 50 µL |
10X rCutSmart Buffer | 5 µL |
DNA | x (1 µg) |
Enzyme 1 | 1 µL |
Enzyme 2 | 1 µL |
Notes
- NEB High-Fidelity (HF) enzymes are ordered whenever possible, they all use rCutSmart reaction buffer and are usually time-saver qualified
- NEB has the option to select your specific enzymes and generate a detailed protocol - Heat inactivation may be recommended if there are additional steps in the workflow without a DNA purification step
References, related resources, and acknowledgments