Features and Benefits
- High fidelity PCR
Proofreading activity with 3' → 5' exonuclease
- High specificity
Better specificity than Taq DNA polymerasde with reduced chance of non-specific products
- Terminal transferase activity
Guaranteed blunt-ended PCR product by blocking terminal transeferase activity
- Reproducibility
Reproducible results with uniform quality products for each batch under the ISO 9001 quality system
Application
- Gene synthesis
- PCR or primer extension requested high fidelity
- Blunt-end PCR cloning or mutagenesis requested high fidelity
Experimental data
Amplication of limiting amounts of template DNA
Figure. Lambda DNA was amplified using 1 units of enzyme in 20 µl reaction volume.
▶ Specifications
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5' to 3' exonuclease activity
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No
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3' to 5' exonuclease activity
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Yes
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3' – A overhang
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No
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Fragment size
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< 20 kb
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▶ Components
• 10X reaction buffer: MgSO4, Tris-HCl, KCl, (NH4)2SO4, Acetylated BSA pH 9.0
• 10 mM dNTPs mix: 2.5 mM of each dNTP
▶ Concentration
250 Units (2.5 U/μl)
▶ Storage conditions
50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, Stablizers, 50 % Glycerol pH 8.2
▶ Storage temperature
-20°C
▶ Unit definition
One unit is defined at the amount of enzyme that will incorporate 10 nmole of dNTP into acid-insoluble material in 30 minutes at 72°C.
▶ Experimental data
Amplication of limiting amounts of template DNA
Figure 1. Human DNA was amplified using 1 units of enzyme in 20 µl reaction volume.
Figure 2. Amplification of fragments ranging from 2 kb to 5 kb from template Lambda DNA 100 pg with 1 units of Pfu DNA Polymerase.
