ExcelTaq™ Hot Start II DNA Polymerase
5U/μl, 500 U x 1
The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction (Fig. 1). The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C (Fig. 2). The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.
The high specificity (Fig. 3) and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA (Fig. 4) or 1 fg of plasmid DNA (Fig. 5). With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications
•Aptamer-based hot start PCR
•Reversible enzyme inactivation
•Omits extra enzyme activation step
•Convenient for room temperature PCR set-up
•High yield and specificity of target amplicons
•Wide range of amplicon length (up to 10 kb)
•High sensitivity (as low as 1 fg of plasmid)
|Hot Start II DNA Polymerase (5 U/μl)
|10X HS Buffer
||2 x 1 ml
50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol
10X HS Buffer
200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM KCl, 100 mM (NH4
,20 mM MgCl2
, 1% Triton X-100
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C
Figure 2. ExcelTaq™ Hot Start II shows absolute amplicon when pre-incubation at 37˚C and 42˚C.
Reactions were subjected to a pre-incubation at 37˚C or 42˚C for 10 minutes before initial denaturation step. General Taq DNA polymerase generated primer dimers and failed to amplify target due to enzyme activated at 37˚C or 42˚C.
Figure 3. ExcelTaq™ Hot Start II DNA Polymerase shows high specificity on amplifying target DNA.
The optimal annealing temperature of GAPDH primer set is 58˚C. Improper annealing temperature set at 52˚C may force primer-dimer formation. ExcelTaq™ Hot Start II DNA Polymerase eliminated primer-dimer and increased amounts of desired product. Taq DNA polymerase reduced target amplification due to primer-dimer formation at improper annealing temperature of 52˚C
Figure 4. ExcelTaq™ Hot Start II DNA Polymerase shows high sensitivity to amplify from low amount of templates.
Each set of PCR reactions contained either 1 pg, 10 pg, or 1 ng of HeLa cell cDNA as templates. ExcelTaq™ Hot Start II DNA Polymerase successfully amplified targets from lower amount of templates, in comparison with hot-start DNA polymerases from other suppliers and general Taq DNA polymerase.
Figure 5. ExcelTaq™ Hot Start II DNA Polymerase amplifies from plasmid template amounts as low as 1 fg.
-20°C for 24 months