Abbexa's DNA Polymerase is a thermostable enzyme possessing 5'-3' DNA polymerase and 3'-5' proofreading exonuclease activities, offering very high yield and high fidelity, even with demanding applications. This DNA polymerase produces blunt-ended amplicons of up to 5 kb in length. This product is supplied with 2 × 2 ml 10X Reaction Buffer containing Mg²⁺, which provides optimal final reaction conditions for most experiments. In order to allow optimization of reaction conditions, 1.2 ml 50 mM MgCl₂ is also provided.

Target	DNA Polymerase
Tested Applications	PCR
Form	Liquid
Storage	Store all components at -20 °C. It is not recommended to store the enzyme at -80 °C as ice crystals may form on the active site, which can affect the enzyme activity. Avoid repeated freeze/thaw cycles.
Validity	Up to 12 months.
Buffer	10X Reaction Buffer: 600 mM Tris-HCl, 60 mM (NH4)2SO4, 100 mM KCl, 20 mM MgSO4, pH 8.3 at 25 °C.
Biological Activity	One unit is defined as the amount of enzyme that incorporates 10 nmol of dNTPs into acid-insoluble form in 30 minutes at 72 °C.
Concentration	2.5 U/µl
Availability	Shipped within 3-7 working days.
Note	THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use	Reaction Components: Component Volume Template Variable, as required Primers (20 µM each) 1 µl 10X Reaction Buffer 5 µl 100 mM dNTP Mix 0.5 µl DNA Polymerase 1 µl Water Variable, up to 50 µl Total Volume 50 µl Thermal Cycling Conditions: Step Number of Cycles Temperature Time per Cycle Initial Denaturation 1 cycle 95-98 °C 3 min Denaturation 25-35 cycles 95-98 °C 15 seconds Annealing 55-60 °C (primer dependent) 15 seconds Extension 72 °C 1.5-2 kb/min Notes: A final concentration of 2 mM MgSO4 is sufficient for amplification of most targets and should only be adjusted if necessary. It is recommended to start with 1 µl (2.5 U) enzyme per 50 µl reaction volume. Forward and reverse primers are generally used at a final concentration of 0.2-0.6 µM each. It is recommended to start with 0.4 µM as the final concentration (i.e. 20 pmol of each primer per 50 µl reaction volume). A primer concentration that is too high can reduce the specificity of priming, resulting in non-specific products. Primers should have a melting temperature (Tm) of approximately 60 °C. The amount of template in the reaction depends mainly on the type of DNA used. For templates with low structural complexity, such as plasmid DNA, it is recommended to use 50 pg-10 ng DNA per 50 µl reaction volume. For eukaryotic genomic DNA, it is recommended to use a starting amount of 200 ng DNA per 50 µl reaction, this can be varied between 5 ng - 500 ng. It is important to avoid using templates re-suspended in EDTA-containing solutions (e.g. TE buffer) since EDTA chelates free Mg2+.

Research Articles on DNA Polymerase

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