Lambda Exonuclease

Description

A highly processive enzyme that acts in the 5´ to 3´ direction, catalyzing the removal of 5´ mononucleotides from duplex DNA. The preferred substrate is 5´-phosphorylated double stranded DNA, although it will also degrade single-stranded and non-phosphorylated substrates at a greatly reduced rate. Lambda Exonuclease is unable to initiate DNA digestion at nicks or gaps (1).

Highlights

Isolated from a recombinant source
Highly processive 5'→3' exonuclease
Supplied in 10X Reaction Buffer

Product Source

A genetic fusion of the E. coli Lambda Exonuclease gene with the gene encoding maltose binding protein (MBP). Following affinity chromatography, Lambda Exonuclease is cleaved from the fusion construct and purified away from MBP.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
Lambda Exonuclease Reaction Buffer-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to produce 10 nmol of acid-soluble deoxyribonucleotide from double-stranded substrate in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X Lambda Exonuclease Reaction Buffer with 1 μg sonicated duplex [3H]-DNA.

Reaction Conditions

1X Lambda Exonuclease Reaction Buffer
Incubate at 37°C

1X Lambda Exonuclease Reaction Buffer:
67 mM Glycine-KOH
2.5 mM MgCl2
50 μg/ml BSA
pH 9.4 @ 25°C

Usage Concentration

5,000 units/ml

Storage Temperature

-20°C

Storage Conditions

25 mM Tris-HCl
50 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 8.0 @ 25°C

Heat Inactivation

75°C for 10 min

Molecular Weight

Theoretical: 28000 daltons

Specific Activity

50,000 units/mg

References

  1. Little, J.W. (1981). Gene Amplif. Anal. 2, 135-145.

FAQs

  1. Can Exonuclease I be used with a double stranded exonuclease to clean up plasmid preparations?
  2. How Does T5 Exonuclease differ from Lambda Exonuclease (NEB# M0262)?
  3. How Does T7 Exonuclease differ from Lambda Exonuclease (NEB# M0262)?
  4. Will Lambda Exonuclease work in CutSmart Buffer?

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Selection Charts

Interactive Tools

Quality Control

Quality Assurance Statement

  • Purified free of contaminating endonucleases and exonucleases.

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Endonuclease Activity (Nicking, Buffer):
    The buffer is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.