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Promega™ Exonuclease III


A 3'→5' exonuclease specific for double-stranded DNA

Manufacturer: promega™  M1815

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Catalog No. PR-M1815

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Description & Specifications

Specifications

Product Name Exonuclease III
Format Liquid
Concentration 150 to 200U/μL
Color Colorless
Inhibitors 50mM NaCl
Source Recombinant E. coli strain
Purity >90%
pH 7.5
Molecular Weight 28000
Includes 10X Reaction Buffer: 660mM Tris HCl (pH 8 at 25°C), 6.6mM MgCl2
Quantity 25,000U
For Use With (Application) Generating nested sets of deletions with double-stranded, linear DNA, preparation of single-stranded DNA
Storage Buffer 20mM Tris HCl (pH 7.5 at 25°C), 1mM DTT, 100mM KCl, 50% Glycerol
Storage Requirements -20°C

The enzyme catalyzes the stepwise removal of mononucleotides starting from a 3'-OH at nicks, blunt ends, recessed ends and 3'-overhangs of less than four bases, yielding nucleoside 5'-phosphates. Exonuclease III will also degrade DNA from 3'-phosphate ends due to its intrinsic 3'-phosphatase activity. In addition, this enzyme has apurinic endonuclease activity and ribonuclease H activity.

  • Flexible—control deletion rate by varying incubation temperature
  • May be inactivated by heating to 75°C for 10 minutes
  • Used in conjuction with S1 nuclease for unidirectional deletion of sequences from the termini of DNA fragments
  • Source: Recombinant E. coli strain
  • Provided with 10X Reaction Buffer (660mM Tris-HCl [pH 8.0 at 25°C], 6.6mM MgCl2)
  • Concentration: 150–200U/μL
  • Unit definition: One unit is defined as the amount of enzyme required to produce 1nmol of acid-soluble nucleotides from double-stranded DNA in 30 minutes at 37°C

Generating nested sets of deletions with double-stranded, linear DNA, preparation of single-stranded DNA