Digests

Protocol

1. Check the NEB (or other supplier) website to confirm enzyme compatibility

2. In a PCR tube add the reaction components according to the table listed below

3. Mix contents gently and spin down using the small benchtop centrifuge

3. 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 Hi

-         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

-         https://nebcloner.neb.com/#!/redigest