FireAmp FastPfu Fly DNA Polymerase High-Fidelity DNA Polymerase

[Product Introduction]

FireAmp FastPfu Fly DNA Polymerase is a hot-start ultra-high-fidelity DNA polymerase designed for rapid PCR. Compared with conventional high-fidelity DNA polymerases,this enzyme features faster amplification speed (≤5 kb fragments can be amplified at an ultra-fast rate of 12 kb/min; >5 kb fragments can be amplified at a high speed of 6 kb/min), higher amplification efficiency, higher yield, higher fidelity, and higher sensitivity. The 2×FireAmp FastPfu Fly Reaction Mix already contains dNTPs. For DNA amplification, only the template, primers, water, and FireAmp FastPfu Fly DNA Polymerase need to be added to adjust the reaction mix to 1× concentration for the reaction. The amplification product is blunt-ended and can be directly used for cloning experiments.

·Fidelity is 108 times that of conventional Taq DNA polymerase.

·Amplification product is blunt-ended and can be directly used for cloning experiments.

·Amplification of genomic DNA fragments (≤15 kb)

·Amplification of plasmid DNA fragments (≤20 kb)

[Specification] 250 units

 

[Features]

· Hot-start, high specificity

· High amplification efficiency

· Rapid, ultra-high fidelity

· High sensitivity

· High yield

[Applications]

· Amplification of complex templates and high GC/AT content templates

· Ultra-high-fidelity rapid PCR, blunt-end cloning, site-directed mutagenesis

· Long-fragment amplification

FG-FG2104 FireAmp FastPfu Fly         [Product Components]

Components
FireAmp FastPfu Fly DNA Polymerase	250 U×1	500 U×1	500 U×6
2×FireAmp FastPfu Fly Reaction Mix	1 ml×3	1.2 ml×5	1.2 ml×30
50 mM MgSO₄	200 μl×1	400 μl×1	1 ml×1
6×DNA Loading Buffer	500 μl×1	1 ml×1	1 ml×2
PCR Stimulant	200 μl×1	400 μl×1	1 ml×1

 

[Recommended PCR System and Conditions (50 μl reaction system)]

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
2×FireAmp FastPfu Fly Reaction Mix	25 μl	1×
FireAmp FastPfu Fly DNA Polymerase	1 μl	2.5 units
Nuclease-free Water	Variable	-
Total volume	50 μl	-

 

[Optimized Parameters (50 μl reaction system)]

Template	Input
Genomic DNA	10-500 ng
Plasmid DNA	1-30 ng
cDNA	1-2 μl cDNA from RT reaction (50-500 ng RNA for RT reaction)

 

[PCR Reaction Protocol]

Number of Cycles	Temperature	Time
1 cycle	98℃	1 min
30-35 cycles	98℃	10 sec
	Tm-5℃	5 sec
	72℃	6 or 12 kb/min *
1 cycle	72℃	1 min

*For fragments ≤ 5 kb, select 12 kb/min; for fragments > 5 kb, select 6 kb/min.

[PCR Stimulant]

PCR Stimulant is used to optimize the amplification of complex templates or high GC/AT content templates. It significantly enhances amplification for Pfu series enzymes. The storage solution concentration is 5×, and the working concentration can be adjusted between 0.5× and 2.0×.

[Precautions]

·It is recommended to add FireAmp FastPfu Fly DNA polymerase to the reaction system as the last step.

·Please thaw the 2×FireAmp FastPfu Fly Reaction Mix completely before use.If a small amount of precipitate forms, dissolve it by heating in a 37°C water bath and mix well before use.