Robust and fast DNA Polymerase with enhanced fidelity
also known as Phusion High-Fidelity DNA Polymerase
| Cat. No. | Amount | Price (EUR) | Buy / Note |
|---|---|---|---|
| PCR-391S | 100 units | 121,70 | Add to Basket/Quote Add to Notepad |
| PCR-391L | 5 x 100 units | 486,70 | 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 70 °C.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid
Concentration: 2.0 units/μl
Description:
Ultra DNA Polymerase is a genetically optimized DNA polymerase for robust, fast and accurate amplification, even with difficult or GC-rich DNA templates. The polymerase is based on Pfu with a fused DNA binding domain.
The polymerase is tolerant against various inhibitors allowing stable amplification with minimized assay optimization.
The enhanced processivity guarantees highly efficient amplification and makes the enzyme the ideal choice for routine applications in analytical or diagnostic assays, cloning and PCR with long or difficult templates.
With a 2x increased extension rate and a 50x increased fidelity compared to Taq, Ultra DNA Polymerase generates improved product yields at high speed without compromising accuracy.
Content:
Ultra DNA Polymerase
2.0 units/μl High Fidelity Polymerase in 20 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5 % Tween-20, 0.5 % Nonidet P-40, 50% (v/v) Glycerol, pH 8.0 (25 °C) and 0.2 mg/ml BSA
Ultra DNA Buffer
5x conc.
PCR Reaction Setup
The PCR procedure below shows appropriate volumes for a single 50 μl reaction. For multiple reactions, prepare a master mix of components common to all and then dispense appropriate volumes into each PCR reaction tube prior to adding template DNA and primers.
Thaw, mix, and briefly centrifuge each component before use.
Add the following components to a microcentrifuge tube:
Recommended 50 μl PCR assay:
| comp. | stock conc. | final conc. | 1 assay @ 20 μl | 1 assay @ 50 μl |
| PCR-grade Water | fill up to 20 μl | fill up to 50 μl | ||
| Ultra DNA Buffer | 5x | 1x | 4 μl | 10 μl |
| dNTP Mix / 10 mM #NU-1006 | 10 mM | 200 μM | 0.4 μl | 1 μl |
| Ultra DNA Polymerase | 2 units/μl | 0.025 units/μl | 0.2 μl | 0.5 μl |
| primer mix or each primer | 10 μM each primer | 200 - 400 nM each primer | 0.4 - 0.8 μl | 1 - 2 μl |
| template /sample DNA | < 10 ng DNA | < 20 ng DNA |
Mix and briefly centrifuge the components.
Recommended cycling conditions:
| initial denaturation | 98 °C | 30 sec | 1x |
| denaturation annealing1) elongation2) | 98 °C 45-68 °C 68-72 °C | 5-10 sec 10-30 sec 15-30 sec/kb | 25-35x |
| final elongation | 72 °C | 5-10 min | 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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