Jun 19, 2020
Version 1
Protocol for Lambda Exonuclease (NEB #M0262) V.1
This protocol is a draft, published without a DOI.
- New England Biolabs1
- 1New England Biolabs
- New England Biolabs (NEB)Tech. support phone: +1(800)632-7799 email: info@neb.com
Protocol Citation: New England Biolabs 2020. Protocol for Lambda Exonuclease (NEB #M0262). protocols.io https://protocols.io/view/protocol-for-lambda-exonuclease-neb-m0262-7r5hm86
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: September 27, 2019
Last Modified: March 29, 2021
Protocol Integer ID: 28189
Keywords: lambda exo
Abstract
Lambda Exonuclease efficiently degrades 5' phosphorylated linear dsDNA from 5' to 3' direction, leaving supercoiled dsDNA intact.*
*Note: For more precise results we recommend titration of the enzyme to the intended substrate.
Materials
MATERIALS
Lambda Exonuclease - 1,000 unitsNew England BiolabsCatalog #M0262S
Safety warnings
Please see SDS (Safety Data Sheet) for hazards and safety warnings.
Set-up the reaction as follows:
| Components | 50 μl REACTION | |
| DNA | up to 5 μg | |
| Lambda Exonuclease Reaction Buffer (10X) | 5 μl (1X) | |
| Lambda Exonuclease | 1 μl (5 units) | |
| Nuclease-free H2O | up to 50 μl |
Incubate at 37 °C for 00:30:00 .
Stop reaction by adding EDTA to 10 millimolar (mM) .
Heat Inactive at 75 °C for 00:10:00 .
To clean up treated samples, we recommend using one of the following steps:
b. Running the reaction on an agarose gel, and then extracting the DNA (we recommend the Monarch Gel Extraction Kit, NEB #T1020), or
c. Performing a phenol/chloroform extraction followed by ethanol precipitation.