Oct 07, 2021
Restriction endonuclease reactions
- 12021 iDEC NEFU_China
- NEFU_China 2021
Protocol Citation: Shuning Guo 2021. Restriction endonuclease reactions. protocols.io https://dx.doi.org/10.17504/protocols.io.byutpwwn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: October 07, 2021
Last Modified: October 07, 2021
Protocol Integer ID: 53875
Keywords: restriction endonuclease reactions
Abstract
This protocol is used to having restriction endonuclease reactions based on endonuclease by NEB.
Materials
Water bath
ddH2O
Endonuclease (NEB)
10 x NEBuffer
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 |
Incubate at 37℃ in water bath for 1 hour.
Stop the reaction by heat inactivation. Temperature and time for inactivation depends on the enzyme.
Use electrophoresis to confirm if correct construct was present.