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Pfu DNA Polymerase
Pfu DNA Polymerase
Place of Origin:
China
Brand:
Shanghai ShineGene Molecular Biotech,Inc
Model:
Supplier:
Shanghai ShineGene Molecular Biotech,Inc.
Price:
Hits:
2486 
Updated:
3/21/2020
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    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.
    Concentration:5U/ul
    Source:
    Pyrococcus furiosus strain Vc1 DSM3638.
    10X Reaction Buffer with MgSO4:
    200mM Tris-HCl (pH 8.8 at 25oC), 100mM KCl, 100mM (NH4)2SO4, 20mM MgSO4, 1.0% Triton X-100 and 1mg/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

    10X Pfu buffer

    5ul

    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.Use dry ice for shipping.
    bio-equip.cn
    ShineGene Bio-Technologies, Inc., located in Shanghai, P.R. China, is a bio-company specialized in providing bio- technical services including Gene Synthesis,Peptide Synthesis,Real Time PCR, Fluorogenic Probe Design,Fluorogenic Probe Label,HRP Enzyme Stabilizer,X-gal,IPTG,ProClin300etc. and supplying reagents such as Hotstart Taq polymerase, dNTPs, qPCR mastermix,one step qPCR kits,First Strand cDNA Synthesis Kits,etc. ShineGene also undertake some OEM or OED orders as well as production task.ShineGene works at the cutting edge of development in PCR, quantitative real-time PCR, transposon and RNA technologies. ShineGenes mission is to provide new tools of excellent quality and extreme purity to the scientific community. Therefore research and development is a continuous process in our company.

    http://www.synthesisgene.com
    http://www.shinegene.org.cn/englsih
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