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You are here:Home » Cloning » Cloning-Enzymes » Exonuclease III

Exonuclease III

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Specifications

Exonuclease III is a 3-5 exonuclease releasing 5 mononucleotides from dsDNA and exhibits AP-specific endonuclease, 3 phosphatase and RNAse H activities.
Catalog #E9008
SourceE.coli cells carrying a cloned E.coli xth gene.
Storage BufferSupplied as a liquid in 50mM Tris-HCl, pH 8.0, 50mM KCl, 1mM DTT, 50% glycerol.
Reaction Buffer (10X) E9008-05Supplied as a liquid in 660mM Tris-HCl, pH 8.0, 6.6mM MgCl2
Unit DefinitionOne unit of Exonuclease III catalyzes the release of 1nmol of acid soluble reaction products from E.coli [3H]-DNA in 30 min at 37°C.
ApplicationsSuitable for use in construction of nested unidirectional deletions of DNA fragments, generation of single-stranded template for DNA dideoxy sequencing and site-directed mutagenesis.
Storage and StabilityMay be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Quality Control
Endodeozyribonuclease Assay
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 50 units of Exonuclease III with 1ug of pUC19 DNA in 50ul of activity assay for 4 hours at 37ºC.
Inhibition and Inactivation
InhibitiorsMetal chelators, p-chloromercuri benzoate (50-90% inhibitory at 0.1mM)
Inactivated by heating at 70°C for 10 min.
Alternate namesSupplied with Reaction Buffer


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