Pfu DNA Polymerase Kit

Pfu DNA Polymerase Kit

Catalog Number	K0566001
Size	250U
Type	PCR Kit
Datasheet

Description

Pfu DNA polymerase isolated from Pyrococcus furiosus, a proofreading DNA polymerase due to 3'→5'exonuclease activity exhibits the lowest error rate of any thermostable DNA polymerase studied. This feature makes Pfu DNA polymerase the ideal choice for a variety of techniques requiring high fidelity DNA synthesis by PCR such as cloning, gene expression, and site-directed mutagenesis. Pfu DNA polymerase is a highly thermostable enzyme, retaining 94-99% of its polymerase activity after 1 hour at 95°C.
To amplify GC-rich regions, denaturation temperature up to 98C can be used to with Pfu DNA polymerase. Pfu DNA polymerase is devoid of terminal deoxynucleotidyltrasferase (TdT) activity, which is characterized by the addition of nontemplate-directed nucleotide(s) at the 3'-end of PCR-generated fragments. Therefore, it generates blunt-ended PCR products. This kit contains all optimized reagents for PCR reaction.

Features

High PCR fidelity with improved yield
Highly thermostable: active more than 4 hrs at 95°C
Source: E.coli, recombinant gene from Pyrococcus furiosus
Lowest error rate
3'→5' exonuclease activity
No extra A addition activity: Generate blunt-ended PCR products
Amplification range:
- Lambda DNA: up to 60Kb
- Human genomic DNA: up to 2Kb
Composition of all optimized reagents for PCR reaction (Pfu DNA Polymerase, Pfu PCR buffer containing MgSO4, dNTP mix, loading dye, nuclease free water and Magic buffer) in separate tubes
A supply of enhancer (Magic buffer) for GC rich template or secondary structures

Applications

High Fidelity PCR experiments
Blunt-end Cloning
Sequencing
Gene expression
Site-directed mutagenesis

Concentration

2.5 units/ul

Unit Definition

One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmol of dNTPs into an acid-insoluble product in 30 minutes at 74°C.

Packing

Component	Cat.No	Packing
Pfu DNA Polymerase	K0566001	250U
10X Pfu PCR Buffer	K0565011	1.5ml
10X dNTP Mix (2.5mM each)	K1756011	0.5ml
10X Loading Buffer	K0561051	0.5ml
Nuclease Free Water	K0561061	0.5ml
4X Magic Buffer	K0561031	0.5ml

Lambda DNA was amplified using 8 different concentrations of Pfu DNA Polymerases.