Thermo Scientific™ Exonuclease VII

Description

• Exonuclease VII is a strict single-strand directed enzyme with 5'→3' and 3'→5' exonuclease activities, making it the only bi-directional exonuclease with single-stranded specificity
• Exonuclease VII has no apparent requirement for divalent cations, as it is fully active in the presence of EDTA
• Initial reaction products are acid insoluble oligonucleotides which are further hydrolyzed into acid soluble form
• The products of limited digestions are small oligomers (dimers to dodecamers)
• Molecular Weight: xseA subunit = 51.8 kDa; xseB subunit = 8.8 kDa\ Optimum Temperature: 37 °C Optimum pH: 8.0 Inactivation: 95 °C for 10 min
• Greater than 95% pure as determined by SDS-PAGE
• Tested for contaminating endonucleases, double-stranded exonucleases, and ribonucleases
• 50 mM Tris-HCl (pH 8.0), 200 mM NaCl, 0.5 mM EDTA, 10 mM DTT, 50% glycerol
• The reaction (50 μL) contains 50 mM Tris-HCl (pH 7.9), 50 mM potassium phosphate (pH 7.6), 8.3 mM EDTA, 10 mM 2-mercaptoethanol, denatured DNA, and enzyme
• One unit is the amount of enzyme required to catalyze the conversion of 1 nmol of nucleotide to acid-soluble nucleotide in 30 min at 37 °C under standard assay conditions
• E
• coli strain containing overproducing clones encoding both the large (xseA) and small (xseB) subunits of Exonuclease VII
• Typical Reaction Conditions (50 μL) 70 mM Tris-HCI, pH 8.0 8 mM EDTA 10 mM 2-mercaptoethanol 1 μg DNA 50 μg/mL0.2 units Exonuclease VII Incubate at 37 °C for 30 min
• Inactivate the enzyme by heating to 95 °C for 10 min
• Note that Exonuclease VII is not inhibited by EDTA
• A typical dilution buffer for use with Exonuclease VII is 50 mM Tris, pH 7.9, 1 mM DTT and 0.5 mg/mL acetylated BSA
• Mapping positions of introns in genomic DNA
• Removal of vector DNA from inserts with poly (dA-T) tails
• Removal of excess PCR primers
• Removal of single-stranded over-hangs produced by restriction endonucleases.