Dephosphorylation of 5´-ends of DNA using rSAP (M0371)
New England Biolabs
Published: 2022-02-21 DOI: 10.17504/protocols.io.bg9kjz4w
Abstract
Protocol for Dephosphorylation of 5´-ends of DNA using rSAP (M0371).
Before start
Steps
1.
* 1 pmol of DNA ends is about 1 µg of a 3 kb plasmid **Scale larger reaction volumes proportionally
Prepare a 20µL reaction as follows:
| A | B |
|---|---|
| DNA | 1 pmol of DNA ends* |
| CutSmart® Buffer (10X) | 2 µl |
| rSAP | 1 unit |
| H2O, purified | to 20 µl** |
Note
Note
2.
Incubate at 37°C for 0h 30m 0s.
3.
Stop reaction by heat-inactivation at 65°C for 0h 5m 0s.
