Fermentas* Endonuclease V, T.maritima (Endo V)

• Active towards abasic sites and urea sites, base pair mismatches, flap and pseudo Y structures, and small insertions/deletions in DNA molecules
• Temperature optimum of activity at 65-70°C

• The absence of other endo-, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests

Source:

• E.coli with a cloned nfi gene of Thermotoga maritima

Molecular Weight:

• 25 kDa monomer
• Definition of Activity Unit:
• One unit of the enzyme converts one µg of supercoiled depurinized plasmid DNA into other topological states in 30 min. at 65°C
• Enzyme activity is assayed in the following mixture: 25mM HEPES-NaOH (pH 7.4), 5mM MgCl₂, 5mM DTT, 2% (v/v) glycerol, 2µg of partially depurinated pUC19 DNA

Storage Buffer:

• The enzyme is supplied in: 20mM HEPES-NaOH (pH 7.4), 5mM DTT, 50mM NaCl, 0.1% (v/v) Triton X-100 and 50% (v/v) glycerol

10X Reaction Buffer:

• 500mM Tris-acetate (pH 7.5), 500mM KCl, 10mM EDTA, 0.5% (v/v) Triton X-100

Inhibition and Inactivation:

• Inhibitors: no specific inhibitors have been described
• Inactivated by heating in boiling water bath for 10 min., preferably in the presence of EDTA

• High-throughput methods for mutation research (3, 4)
• Studies in mutagenesis and DNA repair
• Mismatch cleavage
• Genotyping