Home | Molecular Biology | Restriction & Modifying Enzymes | DNA and RNA Modifying Enzymes | Nucleases

Lambda Exonuclease

Recombinant enzyme Thermal inactivation at 80°C in 15 min

Lambda Exonuclease is a highly processive exodeoxyribonuclease that selectively digests the 5'-phosphorylated strand of double-stranded DNA in 5'-3' direction.
  
Loading...
Thermo Scientific Lambda Exonuclease is a highly processive 5'=>3' exodeoxyribonuclease. It selectively digests the 5'-phosphorylated strand of double-stranded DNA. The enzyme exhibits low activity on single-stranded DNA and non-phosphorylated DNA, and has no activity at nicks and limited activity at gaps in DNA (see​ References 1-2)

Highlights

  • Active in Thermo Scientific PCR buffers

Applications

  • Generating single-stranded PCR products for use in:
    • DNA sequencing (see​ Reference 3)
    • Analysis of DNA single-strand conformation polymorphism (SSCP) (see​ Reference 4)
    • Rolling circle amplification
     
  • Producing single-stranded DNA from double-stranded DNA fragments
  • Cloning of PCR products (see Reference 5)

Note

Use of this enzyme in certain applications may be covered by patents and may require a license.

  
Storage Condition-20 C
HazardousNo
10X Reaction Buffer670 mM glycine-KOH (pH 9.4), 25 mM MgCl2, 0.1% (v/v) Triton X-100
Concentration10 U/µL
Definition of Activity Unit
  • One unit of the enzyme catalyzes the release of 10 nmol of acid soluble reaction products from double-stranded substrate in 30 min at 37ºC.
  • Enzyme activity is assayed in the following mixture: 67 mM glycine-KOH (pH 9.4), 2.5 mM MgCl2, 0.1% (v/v) Triton X-100 and 20 µg/mL sonicated E. coli [3H]-DNA.
InactivationInactivated by heating at 80°C for 15 min.
InhibitionInhibitors: salt at relatively low concentration (0.2 M KCl, 0.1 M NaCl), p-chloromercuribenzoate.
Molecular Weight26 kDa monomer
Quality Control
  • The absence of endodeoxyribonucleases confirmed by appropriate quality test.
  • Functionally tested for generating single-stranded DNA from double-stranded PCR product.
SourceE. coli cells with a cloned exo gene of phage lambda.
Storage BufferThe enzyme is supplied in:
25 mM Tris-HCl (pH 8.0), 0.1 mM EDTA, 1 mM DTT, 50 mM NaCl, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol
Lambda Exonuclease activity

Lambda Exonuclease activity

Lambda Exonuclease activity

Schematic of Lambda Exonuclease activity.


References

  1. J. W. Little, An exonuclease induced by bacteriophage lambda: II, Nature of the enzymatic reaction. J. Biol. Chem. 242, 679-686 (1967).
  2. P. G. Mitsis, J. G. Kwagh, Characterization of the interaction of lambda exonuclease with the ends of DNA. Nucleic Acids Res. 27, 3057-3063 (1999).
  3. R. G. Higuchi, H. Ochman, Production of single-stranded DNA templates by exonuclease digestion following the polymerase chain reaction. Nucleic Acids Res. 17, 5865 (1989).
  4. F. Schwieger, C. C. Tebbe, A new approach to utilize PCR-single-stranded-conformation polymorphism for 16S rRNA gene-based microbial community analysis. Appl. Environ. Microbiol. 64, 4870-4876 (1998).
  5. H. Tseng, DNA cloning without restriction enzyme and ligase. Biotechniques. 27, 1240-1244 (1999).
  6. EPO Patent No 0679190B1.

Protocols and Product Manuals

pdf   Lambda Exonuclease - Product Information