Thermostable DNA polymerase for high accuracy
Thermus species, recombinant, E. coli
| Cat. No. | Amount | Price (EUR) | Buy / Note |
|---|---|---|---|
| PCR-204S | 200 units | 52,10 | Add to Basket/Quote Add to Notepad |
| PCR-204L | 5x 200 units | 208,50 | Add to Basket/Quote Add to Notepad |
For general laboratory use.
Please centrifuge briefly before opening (volume ≤2 ml).
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
Description:
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).
Content:
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 μl | 10x High Fidelity Buffer | green cap |
| 200 μM | each dNTP | - |
| 0.2 - 0.5 μM | each Primer | - |
| 1 - 100 ng | template DNA | - |
| 0.5 μl (1.25 units) | High Fidelity Pol | red cap |
| Fill up to 50 μl | PCR-grade water | - |
| initial denaturation | 94 °C | 2 min | 1x |
| denaturation | 94 °C | 20 sec | 20-30x |
| annealing1) | 50 - 68 °C | 30 sec | 20-30x |
| elongation2) | 68 °C | 1 min/kb | 20-30x |
| final elongation | 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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