Description
Quantity: 10 kU
Unit Size: Each
• The T7 Gene 6 Exonuclease hydrolyzes duplex DNA non-processively in the 5'→3' direction from both 5'-phosphoryl or 5'-hydroxyl nucleotides by liberating oligonucleotides as well as mononucleotides, until about 50% of the DNA is acid soluble. It also degrades nucleotides at the gaps and nicks of double-stranded DNA from the 5'-termini and RNA from RNA:DNA hybrids in the 5'→3' direction.
• The T7 Gene 6 Exonuclease is similar to Lambda Exonuclease in that it catalyzes the stepwise hydrolysis of duplex DNA from the 5' termini liberating 5' mononucleotides. However, unlike Lambda Exonuclease, the enzyme has low processivity and it will remove both 5'-hydroxyl and 5'-phosphoryl termini.
• It also degrades RNA and DNA from RNA:DNA hybrids in the 5'→3' direction but is unable to degrade either double-stranded or single-stranded RNA.
• This enzyme has been used for generating single-stranded DNA templates for sequencing or SNP analysis.
• Molecular Weight: 32 kDa
• Optimum pH: 7.5
• Optimum Temperature: 37 °C
• Inactivation: 75 °C for 10 min or add 2 μL of 0.5M EDTA for a 50 μL reaction volume.
• Heating at 75 °C for 10 min or by adding 2 μL of 0.5M EDTA for a 50 μL reaction volume.
• Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating endonucleases and ribonucleases.
• The reaction mixture (50 μL) contains 50 mM Tris-HCI (pH 8.1), 5 mM MgCl_2, 20 mM KCI, 5 mM 2-mercaptoethanol, double-stranded DNA, and enzyme. Incubation is at 37°C for 15 min.
• One unit is the amount of enzyme required to release 1 nmol of acid soluble nucleotide in 15 min at 37 °C under standard assay conditions.
• 50 units/μL
• E. colistrain containing an overproducing clone of the T7 Gene 6 Exonuclease
• Conversion of 0.5 pmol of λ DNA to single-stranded half molecules by 75 units of enzyme in 30 min at 37°C. Verification by agarose gel electrophoresis.
• Controlled stepwise digestion of double-stranded DNA from the 5' termini
• Generating ssDNA templates for sequencing or SNP analysis
