DNA Polymerase Mix

Abbexa's DNA Polymerase Mix is a convenient ready-to-go 2X reaction mix designed to maximize experiment reproducibility. It contains DNA Polymerase (abx461001), MgCl₂ and ultra-pure dNTPs. The mix is optimized and ready-to-use, simply add water, template and primers. DNA Polymerase Mix (2X) dramatically reduces the time needed to set up reactions, thereby minimizing the risk of contamination. Greater reproducibility is ensured, by a reduction in the number of pipetting steps that can lead to pipetting errors. The 500 rxns size is provided as 10 × 1.25 ml DNA Polymerase Mix, and 1.2 ml 50 mM MgCl₂ solution for optional optimization of reaction conditions.

Target	DNA Polymerase Mix
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	The DNA Polymerase Mix (2X) contains 4 mM Mg2+.
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	2X
Availability	Shipped within 3-7 working days.
Note	This product is for research use only. Not for human consumption, cosmetic, therapeutic or diagnostic use.
Directions for use	Reaction Components: Component Volume Template Variable, as required Primers (20 µM each) 1 µl DNA Polymerase Mix (2X) 25 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: The 2X mix contains 4 mM Mg2+ (final concentration 2 mM Mg2+), which is sufficient for amplification of most targets and should only be adjusted if necessary. 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 Mix

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