Kit of thermostable DNA polymerase for high accuracy, dNTPs and reaction buffer
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
Concentration: 2.5 units/μl
High Fidelity Core Kit contains all reagents required for PCR (except template and primer) in one box combining simple handling with high flexibility. The premium quality polymerase, ultrapure dNTPs and the optimized complete reaction buffer ensure superior amplification results.
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
dNTP Mix (white cap)
10 mM each dNTP (dATP, dCTP, dGTP, dTTP)
High Fidelity Buffer (green cap)
Recommended 50 μl PCR assay:
|5 μl||10x High Fidelity Buffer||green cap|
|1 μl||dNTP Mix||white cap|
|0.2 - 0.5 μM||each Primer||-|
|1 - 100 ng||template DNA||-|
|High Fidelity Pol||red cap|
|Fill up to 50 μl||PCR-grade water||-|
Recommended cycling conditions:
|95 °C||2 min||1x|
|denaturation||95 °C||20 sec||20-30x|
|annealing1)||50 - 68 °C||30 sec||20-30x|
|elongation2)||68 °C||1 min/kb||20-30x|
|68 °C||1 min/kb||1x|
For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.
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