HotStarTaq Plus DNA Polymerase (250)

A QIAGEN hot-start DNA polymerase designed for end-point PCR, this enzyme remains inactive at room temperature so that non-specific DNA synthesis and primer-dimer formation cannot occur before thermal cycling. Activity is triggered by an initial heat-activation step of 5 min at 95°C, after which a typical program uses 25–35 cycles of denaturation (0.5–1 min at 94°C), annealing (0.5–1 min at 50–68°C, approximately 5°C below the primer Tm) and extension (1 min at 72°C, or roughly 1 min per kb for products longer than 1 kb), with a final extension of 10 min at 72°C.

Each 100 µl reaction uses 2.5 units of the polymerase and accepts up to 1 µg of template DNA. The supplied 10x PCR Buffer gives a final MgCl2 concentration of 1.5 mM, which can be increased with additional 25 mM MgCl2 when required. Q-Solution is included to improve amplification of GC-rich templates or those with a high degree of secondary structure, and CoralLoad PCR Buffer is also provided; this buffer incorporates a gel-loading reagent and tracking dyes so that PCR products can be loaded directly onto an agarose gel without separate loading dye. CoralLoad PCR Buffer must not be used in capillary sequencers. A No Template Control is recommended in every run, and the enzyme plus its buffers and reagents should be stored immediately upon receipt at –30 to –15°C in a constant-temperature freezer.

According to QIAGEN, the HotStarTaq Plus product line is being discontinued by November 30, 2023, with the AllTaq PCR kits offered as successors thereafter, subject to stock availability.