Accu DNA Polymerase is purified from an E. coli strain carrying a plasmid with a cloned gene encoding a Pyrococcus sp. DNA polymerase. The enzyme catalyzes the incorporation of nucleotides into duplex DNA in the 5’→3’ direction in the presence of Mg2+ at 70~80°C. Accu DNA Polymerase exhibits 3’→ 5’ exonuclease (proofreading) activity, but has no detectable 5’→ 3’ exonuclease activity.
Applications:
Cloning
Amplification
Mutation Analysis
Accurate polymerization
Mutation Analysis
Accurate polymerization
Components:
Storage Buffer
50 mM Tris-HCl (pH 9.0), 100 mM NaCl, 0.1 mM EDTA, 1% Triton X-100, 5 mM DTT, 50% Glycerol, Stabilizers
10× Reaction Buffer*100 mM KCl, 20 mM MgSO4•7H2O, 200 mM Tris-HCl (pH 8.8), 1% Triton X-100, 100 mM (NH4)2SO4 , 1 mg/ml BSA.
*The reaction buffer is supplied as a 10× concentrate and should be diluted for use.
Storage Buffer
50 mM Tris-HCl (pH 9.0), 100 mM NaCl, 0.1 mM EDTA, 1% Triton X-100, 5 mM DTT, 50% Glycerol, Stabilizers
10× Reaction Buffer*100 mM KCl, 20 mM MgSO4•7H2O, 200 mM Tris-HCl (pH 8.8), 1% Triton X-100, 100 mM (NH4)2SO4 , 1 mg/ml BSA.
*The reaction buffer is supplied as a 10× concentrate and should be diluted for use.
Unit Definition:One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 72°C
Quality Control:Nuclease activity is not detected after incubation of g /Hind III DNA with 5 units of Accu DNA polymerase in 50 μl reaction buffer for 18 hours at 37°C.
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