Exonuclease III is a 3-5 exonuclease releasing 5 mononucleotides from dsDNA and exhibits AP-specific endonuclease, 3 phosphatase and RNAse H activities. Source: E.coli cells carrying a cloned E.coli xth gene. Storage Buffer: Supplied as a liquid in 50mM Tris-HCl, pH 8.0, 50mM KCl, 1mM DTT, 50% glycerol. Reaction Buffer (10X) E9008-05: Supplied as a liquid in 660mM Tris-HCl, pH 8.0, 6.6mM MgCl2 Unit Definition: One unit of Exonuclease III catalyzes the release of 1nmol of acid soluble reaction products from E. coli [3H]-DNA in 30 min at 37C. Applications: Suitable for use in construction of nested unidirectional deletions of DNA fragments, generation of single-stranded template for DNA dideoxy sequencing, site-directed mutagenesis, cloning of PCR products and preparation of strand-specific probes. 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 37C. Inhibition and Inactivation: Inhibitiors: Metal chelators, p-chloromercuri benzoate (50-90% inhibitory at 0.1mM) Inactivated by heating at 70C for 10 min. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Form:
Supplied as a liquid in 50mM Tris-HCl, pH 8.0, 50mM KCl, 1mM DTT, 50% glycerol.
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