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EvaEZ Fluorometric Polymerase Activity Assay Kit

EvaEZ™ Fluorometric Polymerase Activity Assay Kit provides an easy and accurate way to determine the activity of a nucleic acid polymerase without using radioisotopes. It contains EvaGreen® dye together with a primed template, dNTPs, and MgCl2 in a Tris buffer system.

qPCR
Catalog #

29051

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2 x 1 mL

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Product Description

EvaEZ™ Fluorometric Polymerase Activity Assay Kit provides an easy and accurate way to determine the activity of a nucleic acid polymerase without using radioisotopes. It contains EvaGreen® dye together with a primed template, dNTPs, and MgCl2 in a Tris buffer system. In the presence of DNA polymerase activity, the primer will be extended to form a double stranded product that can bind EvaGreen dye, resulting in an increase of fluorescence. The rate of increase of fluorescence is positively correlated to the activity of polymerase. The assay was developed for the measurement of Taq DNA polymerase activity. It can also be used for other DNA polymerases such as Pfu, Vent, Phusion®, Bst, Phi29, MMLV, AMV, SuperScript®, T4 DNA polymerase, T7 DNA polymerase, Klenow, and E. coli DNA polymerase I. The activity assay can be conducted at temperatures from 4°C to 75°C.

References

1. BMC Research Notes 6, 360 (2013).

 

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Support & Faq

EvaGreen® dye and Master Mixes

EvaGreen® dye is spectrally similar to FAM or SYBR® Green I, which means no change in optical settings is required for using an EvaGreen®-based master mix. However, there are several differences between the dyes.

Lower background fluorescence: EvaGreen® dye has less background than SYBR® Green I due to its novel “release-on-demand” DNA-binding mechanism.

Brighter and non-inhibitory:  EvaGreen® is less inhibitory in the PCR reaction than SYBR® Green, permitting the use of saturating dye concentration for maximal signal and better high-resolution DNA melt analysis.

Safer and more environmentally friendly: EvaGreen® dye is the first and only PCR dye to date designed to be environmentally safe.

Dye stability: EvaGreen® dye is very stable both during storage and under PCR conditions. SYBR® Green I, on the other hand, is known to degrade following multiple freeze-thaw cycles and under PCR conditions. Moreover, decomposed SYBR® Green I is reported to be even more inhibitory to PCR.

Digital PCR: EvaGreen® dye is currently the only qPCR dye to be used in droplet digital PCR (ddPCR).

Category: EvaGreen® dye and Master Mixes

← FAQs

Biotium offers three different EvaGreen® dye-based qPCR master mixes:

  • Forget-Me-Not™ qPCR Master Mix
  • Fast EvaGreen® qPCR Master Mix
  • Fast Plus EvaGreen® qPCR Master Mix

All three master mixes contain our high-quality EvaGreen® DNA-binding dye and our fast-activating chemically-modified hotstart enzyme, Cheetah™ Taq.

Forget-Me-Not™ qPCR Master Mix is our newest master mix formulation. For most purposes this is the master mix that we would recommend, as it has many benefits: best sensitivity and signal, blue tracking dyes in the master mix and template to reduce pipetting errors, and low cost. Fast EvaGreen® qPCR Master Mix is a sensitive, general use master mix that does not contain blue tracking dyes. Fast Plus EvaGreen® qPCR Master Mix has a slightly different formulation than the regular Fast EvaGreen® qPCR Master Mix, and was designed for more challenging samples.

Category: EvaGreen® dye and Master Mixes

← FAQs

Both of these qPCR master mixes contain Cheetah™ Taq (our chemically modified hotstart DNA polymerase), buffer and dNTPs.

The Fast EvaGreen® Master Mixes also contain the DNA-binding dye EvaGreen®, and are used for fluorescent dye-based qPCR. The only thing the user needs to supply are primers and template.

The Fast Probe Master Mix  is formulated for conducting real-time PCR using an oligo-based probe, such as a TaqMan™ probe, an MGB probe or an AllGlo™ probe. The user needs to supply primers, probe and template.

Category: EvaGreen® dye and Master Mixes

← FAQs

← FAQs

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Shipping/Shelf-life

Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions. Expedited shipment may be requested at time of checkout. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.