Digests: Difference between revisions
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(updated protocoll to reflect neb enzymes) |
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3. Mix contents gently and spin down using the small benchtop centrifuge | 3. Mix contents gently and spin down using the small benchtop centrifuge | ||
3. Incubate at 37°C for | 3. Incubate at 37°C for 30 min if time-saver qualified, 60 minutes if not | ||
{| class="wikitable" | {| class="wikitable" | ||
Latest revision as of 18:48, 17 January 2025
| 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