<
 
  English | 中文
 
 
 
1
Taq Polymerase for qPCR
Pfu DNA Polymerase
Kod DNA Polymerase
dNTPs
Reverse Transcriptase
MiniRNA Prep
First Strand cDNA Synthesis Kit
Real-Time qPCR Kit
One Step qPCR Kit
DNA Marker
Lab Consumables
Lab Equipments
1

Pfu DNA Polymerase

Catalog #

Pack size

Price()

ZP00202

500U

14.40

ZP00203

1000U

27.20

Description:
    Pfu DNA Polymerase is a thermostable enzyme of approximately 90kDa isolated from Pyrococcus furiosus. The enzyme replicates DNA at 75oC, catalyzing the polymerization of nucleotides into duplex DNA in the 5´-3´ direction in the presence of magnesium. Pfu DNA Polymerase also possesses 3´-5´ exonuclease (proofreading) activity. Base misinsertions that may occur during polymerization are rapidly excised by the proofreading activity of the polymerase. Consequently, Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis. Pfu DNA Polymerase-generated PCR fragments are blunt-ended.Elongation rate is 2Kbp/min.
Concentration:
5U/ul  
Source:

    Pyrococcus furiosus strain Vc1 DSM3638.  
5X Reaction Buffer with MgSO4:
    100mM Tris-HCl (pH 8.8 at 25oC), 50mM KCl, 50mM (NH4)2SO4, 10mM MgSO4, 0.5% Triton X-100 and 0.5mg/ml nuclease-free BSA. 
Features:

    High Fidelity: Pfu DNA Polymerase exhibits the lowest error rate of any thermostable DNA polymerase. 
Applications:
    Pfu DNA Polymerase is recommended for use in PCR, primer extension reactions and other applications that demand high fidelity.  
Unit Definition:

    One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmol of dNTPs into acid-insoluble material in 30 minutes at 75oC.  
Reaction Mixture Set Up

1. Gently vortex and briefly centrifuge all solutions after thawing.

2. Keep solutions on ice.

3. Add to a thin-walled PCR tube, on ice:

Reagent

Quantity, for 50 μl of reaction mixture

Final concentration

5X Pfu buffer

10ul

1X

10mM dNTP

1ul

0.2mM

PrimerI(25pmol/ul)

1ul

0.5pmol/ul

PrimerII(25pmol/ul)

1ul

0.5pmol/ul

Pfu(5U/ul)

0.4ul

2U/50ul

Template DNA

variable

50 pg -1 μg

ddH2O

Up to 50ul

 

4. Gently vortex the sample and briefly centrifuge to collect all drops from walls of tube.

5. If using a thermal cycler without a heated lid, overlay the sample with a half volume of mineral

oil or add an appropriate amount of wax.

6. Place samples in a thermal cycler preheated to 96°C and start PCR.

Recommended thermal cycling conditions:  

Step

Temperature,°C

Time ,min

Number of cycles

Initial denaturation

96

4

1

Denaturation

95

0.5

 

Annealing

50-68

0.5-2

25-35

Extension

72

0.5-4

 

Final Extension

72

5

1

Quality Control Tests:
    PCR (activity), SDS-PAGE (purity), endonuclease/nickase.

Storage:
    Pfu DNA Polymerase in 50mM Tris-HCl (pH 8.2 at 25oC), 0.1mM EDTA, 1mM DTT, 50% glycerol and 0.05% CHAPS should be stored at -20oC.U
se dry ice for shipping. 


Pfu DNA Polymerase manual
If you want to get bulk order price,please contact us.E-mail:master@shinegene.org.cn or shinegene@vip.163.com

Related Link
Peptide Synthesis
Chromas
Make Antibody
Lab Consumables
Gene Synthesis
dNTP

 

Copyright 2003-2015 Shanghai ShineGene Molecular Biotech,Inc. All Rights Reserved.
Floor 2,Building A,328#, Wuhe Road,Minhang District,Shanghai,201109,China
Tel:0086-21-54460832    Fax:0086-21-54460831-13    E-mail:master@shinegene.org.cn