Abx071019-250U | FastPfu DNA Polymerase Enzyme (with 2.5 mM dNTPs)

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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 for up to 2 years. Avoid repeated freeze/thaw cycles.

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 human consumption, cosmetic, therapeutic or diagnostic use.

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
	T_M - 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.

Research Articles on FastPfu DNA Polymerase

FastPfu DNA Polymerase Enzyme (with 2.5 mM dNTPs)

Référence abx071019-250U

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