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.
EvaEZ™ Polymerase Activity Mix is provided at 2X concentration with 5 mM MgCl2 (final Mg2+ concentration in assay is 2.5 mM). Magnesium-free EvaEZ™ Polymerase Activity Mix (29051MF) is available on request for customers who wish to control the magnesium concentration in the assay. Contact customer service at firstname.lastname@example.org to inquire about availability.
Phusion and SuperScript are registered trademarks of Thermo Fisher Scientific.
1. BMC Research Notes 6, 360 (2013).