Restriction endonuclease reactions
Shuning Guo
Published: 2021-10-07 DOI: 10.17504/protocols.io.byutpwwn
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Abstract
This protocol is used to having restriction endonuclease reactions based on endonuclease by NEB.
Steps
1.
Add the following reagents to a PCR tube (e.g. 50μl) on ice.
| A | B |
|---|---|
| Restriction Enzyme | 10 units is sufficient, generally 1 µl is used |
| DNA | 1μl |
| 10 x NEBuffer | 5μl |
| ddH2O | To 50μl |
2.
Incubate at 37℃ in water bath for 1 hour.
3.
Stop the reaction by heat inactivation. Temperature and time for inactivation depends on the enzyme.
4.
Use electrophoresis to confirm if correct construct was present.
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