Jun 19, 2020
Version 1
Protocol for Exonuclease VII (NEB #M0379) 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 Exonuclease VII (NEB #M0379). protocols.io https://protocols.io/view/protocol-for-exonuclease-vii-neb-m0379-7sahnae
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: 28194
Keywords: exo VII
Abstract
Exonuclease VII efficiently cleaves single-stranded DNA (ssDNA) from both 5´→3´ and 3´→5´ direction. This enzyme is not active on linear or circular dsDNA
Materials
MATERIALS
Exonuclease VIINew England BiolabsCatalog #M0379
Safety warnings
Please see SDS (Safety Data Sheet) for hazards and safety warnings.
Set-up the reaction as follows:
| COMPONENTS | Reaction Volumes | |
| PCR Product | 5 μl | |
| Exonuclease VII | 2 μl (20 units) |
Incubate the mix at 37 °C for 00:30:00 .
Heat inactivate 95 °C for 00:10:00 prior to Sanger DNA sequencing.
For other downstream molecular cloning applications column cleanup is recommended.
Note
Note: See Exo-CIP™ Rapid PCR Cleanup Kit (NEB #E1050) for Sanger Sequencing, SNP detection, or library preparation for NGS.
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.