Exonuclease III

Kód produktu: EN0191 Kód výrobce: EN0191 Kód dodavatele: {723C1BCB-1807-4C3C-86E0-9F1C2300A0D7} Výrobce: UAB Fermentas
842,16 Kč
696,00 Kč bez DPH
Skladem
4000 Units
Exonuclease III is a 3'–>5' exonuclease specific for double-stranded DNA or DNA-RNA hybrids. Exhibits also 3' phosphatase and purinic/apyrimidinic-endonuclease activities.
Zobraz detailní popis

Detailní popis

Thermo Scientific Exonuclease III (ExoIII) exhibits four catalytic activities (see Reference1). The 3'=>5' exodeoxyribonuclease activity of ExoIII is specific for double-stranded DNA. ExoIII degrades dsDNA from blunt ends, 5'-overhangs or nicks, releases 5'-mononucleotides from the 3'-ends of DNA strands and produces stretches of single-stranded DNA. It is not active on 3'-overhang ends of DNA that are at least four-bases long and do not carry a 3'-terminal C-residue (see Reference 2) on single-stranded DNA, or on phosphorothioate-linked nucleotides.

ExoIII 3'-phosphatase activity removes the 3'-terminal phosphate, generating a 3'-OH group. ExoIII Rnase H activity exonucleolytically degrades the RNA strand in RNA-DNA hybrids. ExpOOO apurinic/apyrimidinic-endonuclease activity cleaves phosphodiester bonds at apurinic or apyrimidinic sites to produce 5'-termini that are base-free deoxyribose 5'-phosphate residues.

Highlights

  • Active in restriction enzyme buffers

Applications

  • Creation of unidirectional deletions in DNA fragments in conjunction with S1 Nuclease (see References 2, 3)
  • Generation of a single-stranded template for dideoxy-sequencing of DNA (see Reference 4)
  • Site-directed mutagenesis (see Reference 5)
  • Cloning of PCR products (see Reference 6)
  • Preparation of strand-specific probes

Note

The rate of DNA digestion by ExoIII depends upon temperature, salt concentration, and the molar ratio of DNA to enzyme in the reaction mixture (see References 4, 8). Optimal reaction conditions should be determined experimentally.

10X Reaction Buffer 660 mM Tris-HCl (pH 8.0 at 30°C), 6.6 mM MgCl2.
CategoryName
Definition of Activity Unit
  • One unit of the enzyme catalyzes the release of 1 nmol of acid soluble reaction products from E. coli [3H]-DNA in 30 min at 37°C.
  • Enzyme activity is assayed in the following mixture: 50 mM Tris-HCl (pH 8.0), 5 mM MgCl2, 1 mM DTT, and 0.05 mM sonicated E. coli [3H]-DNA.
Hazardous No
Hazardous: No
Inactivation Inactivated by heating at 70°C for 10 min.
Inhibition Inhibitors: metal chelators, p-chloromercuri benzoate (50-90% inhibitory at 0.1 mM) (see Reference 7).
Molecular Weight 31 kDa monomer.
Quality Control
  • The absence of endodeoxyribonucleases confirmed by appropriate quality test.
  • Functionally tested for creation of unidirectional deletions in DNA fragments.
Shelf Life:
Shipping Condition:
Shipping Information
Source E. coli cells with a cloned E. colixth gene.
SpecificationName
SpecificationValue
Storage Buffer The enzyme is supplied in:
50 mM Tris-HCl (pH 8.0), 50 mM KCl, 1 mM DTT and 50% (v/v) glycerol.
Storage Condition -20 C
Storage Condition: -20 C

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