Abbexa's Heat-Activated DNA Polymerase with Red Dye Mix is a complete ready-to-use 2X reaction mix containing a heat-activated thermostable enzyme with improved specificity as compared to standard polymerases, by eliminating all non-specific priming and the formation of primer-dimers. It is inactive at room temperature, allowing for reaction set-up at room temperature, and therefore requires activation by heat treatment for 10 minutes prior to PCR cycling. It also contains an additional inert red dye that permits easy visualization and direct loading onto a gel. No loading buffer is required as the mix is of sufficiently high density to sink to the bottom of the gel. The mix has been developed to perform PCR assays of many common genomic and cDNA templates; simply add water, template and primers. It dramatically reduces the time required to set up reactions, thereby minimizing the risk of contamination. Greater reproducibility is ensured, by reducing the number of pipetting steps that can lead to errors. The 500 rxns size is provided as 10 × 1.25 ml Heat-Activated DNA Polymerase with Red Dye Mix, and 1.2 ml 50 mM MgCl2 solution for optional optimization of reaction conditions.
Target
Heat-Activated DNA Polymerase with Red Dye Mix
Tested Applications
PCR
Form
Liquid
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.
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.
Shelf Life
Up to 12 months.
Buffer
The exact formulation is proprietary.
Concentration
2X
Availability
Shipped within 10-20 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 and Primers
Variable, as required
Heat-Activated DNA Polymerase with Red Dye Mix
25 µl
Water
Variable, up to 50 µl
Total Volume
50 µl
Thermal Cycling Conditions:
Activation: pre-heat at 95 °C for 10 minutes
Denaturation: 94-96 °C
Annealing: dependent on primer Tm
Extension: 72 °C, allowing 15-30 seconds per kb
Notes:
The optimal conditions will vary from reaction to reaction and are dependent on the system used. Each parameter needs to be adjusted by the end user and some optimization may be required.
Pre-incubate at 95 °C for 10 minutes to activate the enzyme. Subsequently, the reaction can be treated according to the end user's existing protocol.
If extension time exceeds 2.5 minutes, a maximum of 30 cycles should be used. Increasing the number of cycles may lead to smearing when run on an agarose gel.
The 2X mix contains a final concentration of 3 mM MgCl2 in a 50 µl reaction volume, which is sufficient for amplification of most targets. The provided MgCl2 solution can be used to adjust the Mg2+ concentration if required.
For optimal resolution of PCR products, it is recommended to use Tris-Acetate EDTA (TAE) buffer for gel preparation and electrophoresis.
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