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Pfu DNA Polymerase (native)

Kód produktu: EP0572 Kód výrobce: EP0572/F530L Kód dodavatele: {8F29F6AC-BC3F-4615-8B40-1E71B7AEDBAF} Výrobce: UAB Fermentas
10 599,60 Kč
8 760,00 Kč bez DPH
do týdne
500 Units

Highly thermostable DNA polymerase for PCR applications requiring high fidelity.

Zobraz detailní popis

Detailní popis

Thermo Scientific Pfu DNA Polymerase is a highly thermostable DNA polymerase from the hyperthermophilic archaeum Pyrococcus furiosus. The enzyme catalyzes the template-dependent polymerization of nucleotides into duplex DNA in the 5’→3’ direction. Pfu DNA Polymerase also exhibits 3’→5’ exonuclease (proofreading) activity that enables the polymerase to correct nucleotide incorporation errors. It has no 5’→3’ exonuclease activity.

Highlights

  • Eight times more accurate than Taq DNA polymerase
  • Highly thermostable – remains 95% active after 2 hours incubation at 95°C
  • Generates blunt-end PCR products
  • Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides)
  • Increased PCR yields with native Pfu DNA Polymerase

Applications

  • High fidelity PCR
  • Generation of PCR products for cloning and expression
  • RT-PCR for cDNA cloning and expression
  • Blunt-end PCR cloning (see Reference 1)

Includes

  • Pfu DNA Polymerase (native or recombinant, 2.5 U/µL)
  • 10X Pfu Buffer with MgSO4
  • 10X Pfu Buffer
  • 25 mM MgSO4

Note

The error rate of Pfu DNA Polymerase in PCR is 2.6 x 10-6 errors per nt per cycle as determined by a modified method described by Lundberg et al. (see Reference 2). Accordingly, the accuracy of PCR is 3.8 x 105.  Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs. The Pfu DNA Polymerase has no detectable reverse transcriptase activity. dUTP, dITP, and primers containing these nucleotides should not be used in PCR with Pfu DNA Polymerase because the binding of this enzyme to DNA templates with uracil and hypoxanthine stalls DNA synthesis (see References 3,4).

10X Pfu Buffer 200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM (NH4)2SO4, 100 M KCl, 1% (v/v) Triton X-100, 1 mg/mL BSA.
10X Pfu Buffer with 20 mM MgSO4 200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM (NH4)2SO4, 100 mM KCl, 1 mg/mL BSA, 1% (v/v) Triton X-100, 20 mM MgSO4.
Definition of Activity Unit
  • One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 minutes at 72°C.
  • Enzyme activity is assayed in the following mixture: 20 mM Tris-HCl (pH 8.8 at 25°C), 2 mM MgSO4, 10 mM (NH4)2SO4, 10 mM KCl, 0.1 mg/mL BSA, 0.1% (v/v) Triton X-100, 0.75 mM activated calf thymus DNA, 0.2 mM of each dNTP, 0.4 MBq/mL [3H]-dTTP.
Hazardous No
Inactivation Inactivated by phenol/chloroform extraction.
Molecular Weight 90 kDa monomer
Quality Control
  • The absence of endo- and exodeoxyribonucleases confirmed by appropriate quality tests.
  • Functionally tested in PCR.
Source Pfu DNA Polymerase (native) Pyrococcus furiosusPfu DNA Polymerase (recombinant) E.coli with a cloned pol gene from Pyrococcus furiosus.
Storage Buffer 20 mM Tris-HCl (pH 8.2), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.1% (v/v) Nonidet P40, 0.1% (v/v) Tween 20, and 50% (v/v) glycerol.
Storage Condition -20 C

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