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High Fidelity Polymerase

Thermostable DNA polymerase for high accuracy

Thermus species, recombinant, E. coli

Cat. No. Amount Price (EUR) Buy / Note
PCR-204S 100 units70,30 Add to Basket/Quote Add to Notepad
PCR-204L 500 units281,90 Add to Basket/Quote Add to Notepad

For general laboratory use.

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

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
avoid freeze/thaw cycles

Shelf Life: 12 months

Form: liquid

Concentration: 2.5 units/μl

High Fidelity Pol is based on a blend of Taq DNA polymerase and a proofreading enzyme specially designed for highly accurate and efficient amplification. It shows excellent results with extremely long (up to 30 kb), GC-rich or other difficult templates.
The enzyme blend includes a highly processive 5'→3' DNA polymerase and possesses a 5'→3' polymerization-dependent exonuclease replacement activity. Its inherent 3'→5' exonuclease proofreading activity results in a greatly increased fidelity of DNA synthesis compared to Taq polymerase.
The enzyme is highly purified and free of bacterial DNA.

Fidelity of the enzyme:
High Fidelity Pol is characterized by a 4-fold higher fidelity compared to Taq polymerase.
ERHigh Fidelity Pol = 3.4 x 10-6
The error rate (ER) of a PCR reaction is calculated using the equation ER = MF/(bp x d), where MF is the mutation frequency, bp is the number of base pairs of the fragment and d is the number of doublings
(2d = amount of product / amount of template).

High Fidelity Pol (red cap)
2.5 units/μl High Fidelity Polymerase in storage buffer

High Fidelity Buffer (green cap)
10x conc.

Recommended 50 μl PCR assay:

5 μl10x High Fidelity Buffergreen cap
200 μMeach dNTP-
0.2 - 0.5 μMeach Primer-
1 - 100 ngtemplate DNA-
0.5 μl
(1.25 units)
High Fidelity Polred cap
Fill up to 50 μlPCR-grade water-

Please note that it is essential to add the polymerase as last component.
Recommended cycling conditions:
94 °C2 min1x
denaturation94 °C20 sec20-30x
annealing1)50 - 68 °C30 sec20-30x
elongation2)68 °C1 min/kb20-30x
68 °C1 min/kb1x

1)The annealing temperature depends on the melting temperature of the primers used.
2)The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.

For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.

Related products:

  • Ready-to-Use Mixes / direct gel loading
  • Ready-to-Use Mixes
  • Thermophilic Polymerases
  • Deoxynucleotides (dNTPs)
  • Supplements
  • Primers and Oligonucleotides
  • DNA Ladders

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