FastPfu DNA Polymerase is a fast, high fidelity and high processivity hot start DNA polymerase with an extension rate of 4 kb/min. Amplification of genomic DNA fragment up to 15 kb. Amplification of plasmid DNA fragment up to 20 kb. Contents:
Component
250 U
500 U
3 kU
FastPfu DNA Polymerase
1 × 250 U
1 × 500 U
6 × 500 U
5X FastPfu Buffer
1.2 ml
2 × 1.2 ml
12 × 1.2 ml
2.5 mM dNTPs
500 µl
1 ml
6 × 1 ml
50 mM MgSO4
200 µl
400 µl
1 ml
PCR Stimulant
200 µl
400 µl
1 ml
6X DNA Loading Buffer
500 µl
1 ml
2 × 1 ml
Target
FastPfu DNA Polymerase
Tested Applications
PCR
Conjugation
Unconjugated
Storage
Store at -20°C. Avoid repeated freeze/thaw cycles.
Shelf Life
24 months.
Buffer
FastPfu DNA Polymerase: Contains 50 mM Tris-HCl (pH 8.2), 0.1 mM EDTA, 1 mM DTT, Stabilizers, 50% glycerol. FastPfu Buffer: Contains 100 mM Tris-SO4 (pH 9.2), 200 mM KCl, 10 mM MgSO4, 10% glycerol.
Concentration
2.5 U/µl
Availability
Shipped within 10-20 working days.
Note
THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use
Reaction Components:
Component
Volume
Final Concentration
Template
Variable
as required
Forward Primer (10 µM)
1 µl
0.2 µM
Reverse Primer (10 µM)
1 µl
0.2 µM
5X FastPfu Buffer
10 µl
1X
2.5 mM dNTPs
4 µl
0.2 mM
FastPfu DNA Polymerase
1 µl
2.5 U
Nuclease-Free ddH2O
Variable
N/A
Total Volume
50 µl
N/A
Suggested Reaction Conditions:
Template
Quantity
Genomic DNA
10-100 ng Genomic DNA
Plasmid DNA
1-30 ng Plasmid DNA
cDNA
1-2 µl cDNA from RT reaction (50-500 ng RNA for RT reaction)
Thermal Cycling Conditions:
Number of Cycles
Temperature
Time
1 cycle
95 °C
2 min
30-35 cycles
95 °C
20 seconds
TM - 5 °C
20 seconds
72 °C
6 kb/min
1 cycle
72 °C
5 min
Notes:
The PCR Stimulant can be used to optimize the amplification of complex templates or high GC/AT templates. The recommended working concentration is 0.5X - 2X (concentration of the provided stock solution is 5X).
It is recommended to add the FastPfu DNA Polymerase last in the reaction system.
For GC-rich templates, the recommended denaturation temperature is 98 °C.
If precipitates in the FastPfu Buffer are observed, warm to 37 °C in a water bath and mix before use.
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