Recombinant Nucleases

Recombinant Exonuclease III-T: Engineered exonuclease III with improved temperature resistance.

Enzyme useful for nested unidirectional deletions, generation of a single-stranded templates, site-directed mutagenesis and other cloning tasks.

Exonuclease III (ExoIII) of E. coli is a 31 kD monomeric, globular protein combining multiple catalytic activities in one active site. It acts on doublestranded (ds) DNA as a 3'-5' exonuclease, a 3'- phosphomonoesterase, an apurinic/apyrimidimic (AP) sites specific endonuclease and an exonucleolytic ribonuclease H. Of particular interest for molecular biological methods is its exonucleolytic activity, removing 5'-mononucleotides from the 3'-hydroxyl ends of ds DNA, leaving protruding 5'-termini. Its catalytic rate can be adjusted by temperature and NaCl concentration and thus allows for the generation of single-stranded DNA templates for sequencing or recombination methods.

20μl (>10U/μl)

Applications

• Construction of nested unidirectional deletions of DNA fragments in combination with nuclease S1
  
• Generation of a single-stranded template for dideoxy sequencing of DNA
  
• Site-directed mutagenesis
  
• Cloning of PCR products
  
• In vitro recombination