Page 92 - 2022-23 BIOFACT Catalogue

BioFACT™

Pfu DNA Polymerase

High Fidelity / Ampli cation of Blunt-End Fragment

BioFACT™ Pfu DNA Polymerase is recombinant enzyme derived from Pyrococcus furiosus and expressed in E.coli. It has superior ther-
mostability and possesses 3' to 5' exonuclease proofreading activity which results in PCR fragments with fewer errors. PCR fragments
generated by BioFACT™ Pfu DNA Polymerase are blunt-ended and it can be available for blunt end cloning using All in One™ PCR Cloning
Kit without additional A-tailing step.

Feature
• Source : Pyrococcus furiosus • A-tailing : No
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• 5' → 3' exonuclease activity : No • Error rate : 1-2 bp error / 10 bp
• 3' → 5' exonuclease activity ( delity) : Yes • 12-fold higher PCR delity over Taq DNA Polymerases
• Ampli cation size : < 3 kb PCR

Application
• High delity PCR • Blunt end form cloning
• Site-directed mutagenesis

QC DATA
- Comparison of amplification efficiency according to various sizes of template between BioFACT™ Pfu DNA Polymerase and
other companies' Pfu DNA Polymerases

BioFACT™ Pfu Company A Company B
M 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7

Template : Human genomic DNA, Corynebacterium Ch's phage λ DNA
PCR Conditions : AT 68℃ (3, 4 : 62℃) 30 cycles
M : 1kb DNA Ladder
1 : 0.5 kb 2 : 1 kb 3 : 1.5 kb 4 : 3 kb 5 : 4 kb 6 : 6 kb 7 : 10 kb

- Comparison of fidelity for various PCR enzymes

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DNA polymerase Average Mutation Frequency* Average Error Rate** Relative Fidelity
(%±SD)
±SD)
(10 -6
Enzyme Accuracy (x10 5 ) 6 4 Lamp Taq 4.03±1.43 23.0±19.0 10x
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Taq
1x
2.31±2.2
0.45±0.73
Pfu
2 Lamp Pfu 0.16±0.05 0.96±0.4 24x
1.22±0.9
0.21±0.07
19x
0 • The average of five independent rpsL Fidelity Assays is shown.
Taq Lamp Taq Pfu Lamp Pfu
*Mutation Frequencies were calculated using the equation : MF = (Apr Smr colonies) / [Apr colonies × dilution factor × 100%].
Error rate = mutation / bp / duplication **Error Rates were calculated using the equation : ER = Mutation Frequency / (Template Doublings × 130 bp).
• Template Doublings were determined using the equation : TD = (amount of PCR product) / (amount of starting product).
• Template Doublings for all three polymerases ranged from 11 to 12 for 25 PCR cycles, and 6 to 7 for 15 cycles.
92 BIOFACT CATALOGUE