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HyFD Fusion DNA Polymerase
Description
Fusion DNA polymerase is a recombinant 95 kDa DNA
polymerase obtained by expression of the modified Pyrococcus
furiosus DNA polymerase together with Sulfolobus solfataricus
processivity-enhancing domain in E. coli. This thermostable
enzyme exhibits a strong 5’ to 3’ polymerase activity and 3’ to 5’
exonuclease (proofreading) activity, but no 5’ to 3’ exonuclease
activity.
Ordering Information
FU-01
E-Bio Fusion DNA polymerase
500 units
Supplied with 10X Reaction Buffer (1.25 ml/vial x 2 vials)
incorporate 10 nmol of dNTPs into acid-insoluble material in 30
minutes at 74°C
Unit assay conditions
25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM
DTT, 0.5 mg/ml activated salmon sperm DNA, 0.2 mM dATP,
dCTP, dGTP, dTTP
Storage and Dilution Buffer
50 mM Tris-HCl, pH 8.0 (at 25°C), 1 mM EDTA, 5 mM DTT, 100
mM NaCl, 1% Triton X-100, 50% (v/v) glycerol and stabilizers.
Concentration
2 units/μl
10X Reaction Buffer
750 mM KCl, 100 mM Tris-HCl, pH 8.8, 35 mM MgCl2 and 1%
Triton X-100
Unit Definition
One unit of enzyme is defined as the amount of enzyme that will
Storage Conditions
Store Fusion DNA polymerase at -20°C in a constant freezer
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Example of Reaction Composition
Volume/Reaction
Final Concentration 1X
10X Reaction Buffer
5 μl
1X
dNTP Mix (2.5 mM)
4 μl
200 μM of each dNTP
Forward Primer (10 μM)
Variable
0.1–0.5 μM
Reverse Primer (10 μM)
Variable
0.1–0.5 μM
0.5 μl
1.0 units/reaction
Distilled water (PCR Grade)
Variable
-
Template DNA
Variable
≤1 μg/reaction
Total Volume
50 μl
DNA Polymerase (2 U/μl)
Note: If smaller or larger reaction volumes are used, please reduce or increase the amount of each component accordingly.
Example of Thermal Cycler Conditions
Temp.
Time
Number of Cycle
Initial denaturation
98°C
3
1 cycle
Denaturation
98°C
0.5–1 min
Annealing
50–68°C
0.5–1 min
Extension
72°C
0.5 min/1kb
Final extension
72°C
10 min
25 to 40 cycles
1 cycle
Note: For PCR products longer than 1 kb, use an extension rate at approximately 1 kb/min
For Research Use Only
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