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Molecular Biology Tools

Products >  Molecular_Biology_Tools >  Modifying_Enzymes >  Nucleases >  Exonuclease_III

Exonuclease III

Exonuclease III is a 3' to 5' exonuclease that removes mononucleotides from the 3´-hydroxyl termini of double-stranded DNA or DNA-RNA hybrids. Because a set number of mononucleotides are generated during each degradation, double-stranded DNA degradation is progressive. Exonuclease III will degrade double-stranded DNA containing a blunt end, a 5'-overhang or a nick, but it will not degrade DNA with a 3'-overhang containing 4 or more bases. This 3' to 5' exonuclease can be inactivated by heating it at 65°C for 5 minutes. Exonuclease III is supplied in a buffer of 25 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.5 mM DTT and 50% glycerol.

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  • Preparation of single-stranded DNA for dideoxy sequencing
  • Production of nested deletions in double-stranded DNA
  • DNA-protein interaction analysis 
  • Site-directed mutagenesis


Recombinant E. coli


–20°C (-80°C when stored for more than 6 months)


Supplied with 10X Reaction Buffer [500 mM Tris-HCl (pH 8.0), 50 mM MgCl2, 100 mM DTT]


  1. Wu, C. (1985) Nature 317:84-87.
  2. Guo, L.H. & Wu, R. (1983) Methods Enzymol. 100:60-96.

Unit Definition

One unit is defined as the amount of enzyme required to produce 1 nmol of acid-soluble product in 30 minutes at 37°C and pH 8.0 using restriction-digested calf thymus DNA as the substrate.


200 U/µl

Cat. # Product Contents Size Price Units Select
2170A Exonuclease III 5,000 Units $50.00
2170B Exonuclease III 25,000 Units $200.00



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