TopTaq DNA Polymerase
For highly reliable end-point PCR applications with unrivaled ease of use
TopTaq DNA Polymerase combines a number of convenient and unique features to provide unrivaled ease of use and ensure high DNA yields in a wide range of PCR applications. The unique TopTaq Stabilizer included in the enzyme storage buffer allows storage of TopTaq DNA Polymerase at 4°C and reaction setup at room temperature without time-consuming thawing of reagents. CoralLoad Concentrate, which contains two gel-tracking dyes for easier pipetting visualization and immediate gel-loading of PCR products, is also provided. High yields of PCR products are achieved, even when amplifying a range of different sized products using the same Mg2+ concentrations and annealing temperatures.
TopTaq DNA Polymerase is used for standard and specialized end-point PCR applications. TopTaq DNA Polymerase is the first PCR kit that can be stored at 4°C. TopTaq Stabilizer (contained in the enzyme storage buffer) binds to TopTaq DNA Polymerase, preventing polymerase inactivation during long-term storage. During the initial denaturation step, the TopTaq Stabilizer dissociates from the polymerase without compromising polymerase activity. TopTaq DNA Polymerase outperformed DNA polymerases tested from other suppliers and ensures high yields of specific products, even when using low template amounts, and independent of fragment size (see figure "High PCR product yields without optimization).
TopTaq DNA Polymerase is a recombinant 94 kDa thermostable DNA Polymerase. TopTaq DNA Polymerase has been developed by QIAGEN to provide highly reliable end-point PCR with unrivaled ease of use. Until the development of the TopTaq PCR system, all PCR enzymes required storage at –20°C; however, due to the unique proprietary TopTaq Stabilizer contained in the enzyme storage buffer, TopTaq DNA Polymerase is the first PCR kit that can be stored at 4°C.
TopTaq DNA Polymerase, in combination with the innovative TopTaq PCR Buffer, facilitates amplification of specific PCR products by providing a high ratio of specific-to-nonspecific primer binding. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers (see figure "Increased specificity of primer annealing"). The need to optimize PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced, or often not required. Thus, TopTaq DNA Polymerase provides the same reliability and specificity as Taq DNA Polymerase and additionally offers improved performance in some PCR systems — resulting in higher yields of PCR products (see figure "High yields of PCR product without optimization").
TopTaq Stabilizer binds to TopTaq DNA Polymerase at 4°C and room temperature, preventing polymerase denaturation during long-term storage. Template DNA and primers are also prevented from binding to the polymerase at low temperature. During the initial denaturation step, the TopTaq Stabilizer dissociates from the polymerase without compromising polymerase activity (see figure "Stabilization at 4°C").
For enhanced convenience, TopTaq DNA Polymerase is supplied with CoralLoad Concentrate. CoralLoad Concentrate contains a gel-loading reagent and two gel-tracking dyes. This unique reagent improves pipetting visibility and facilitates estimation of DNA migration distance and optimization of agarose gel run time. When using CoralLoad Concentrate, PCR products can be directly loaded onto an agarose gel without prior addition of loading buffer (see figure "Convenient CoralLoad Concentrate").
Optimized protocol and buffer
With its balanced potassium and ammonium ions, the unique PCR Buffer, used in combination with TopTaq DNA Polymerase, promotes specific primer–template annealing and simultaneously reduces nonspecific annealing. Maximum yields of specific products are obtained even when using low template amounts.The unique buffer formulation and single preoptimized protocol eliminate the need for optimization of experimental parameters for individual-primer template systems, saving time and costs. High yields of specific amplification product are achieved, even when amplifying a range of fragments of different sizes using the same Mg2+ concentration and annealing temperature (see figure "High PCR product yields without optimization").
This innovative PCR additive facilitates amplification of difficult templates by modifying the melting behavior of DNA. Use of this unique reagent will often enable or improve suboptimal PCR. Unlike DMSO and other PCR additives, Q-Solution is used at a defined working concentration with any primer-template system and is not toxic.
Due to the unique proprietary TopTaq Stabilizer contained in the enzyme storage buffer, TopTaq DNA Polymerase is the first PCR kit that can be stored routinely at 4°C. During the initial denaturation step, the TopTaq Stabilizer freely dissociates from the polymerase without compromising polymerase activity. This added convenience allows immediate reaction setup without the use of ice — saving the time usually required for reagent thawing (see flowchart "TopTaq procedure"). All components can be combined at room temperature eliminating the need for working on ice. The unique buffer formulation and a single preoptimized protocol eliminate the need for optimization of experimental parameters for individual primer–template systems.
TopTaq DNA Polymerase is highly suited to all end-point PCR applications.
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