HotStarTaq Plus Master Mix Kit (1000)

QIAGEN supplies this kit as a ready-to-use mixture of HotStarTaq Plus DNA Polymerase, QIAGEN PCR Buffer, MgCl2, and dNTPs, designed for fast and highly specific amplification across a range of PCR applications. The polymerase is a modified form of QIAGEN Taq DNA Polymerase provided in an inactive state and activated by a 5-minute incubation at 95°C, which prevents extension of nonspecifically annealed primers and primer–dimers formed during PCR setup and the initial cycle.

The included QIAGEN PCR Buffer combines KCl and (NH4)2SO4 to maintain a high ratio of specific-to-nonspecific primer binding, reducing the need to optimize annealing temperature or Mg2+ concentration. CoralLoad Concentrate is also included, providing a gel-loading reagent and two gel-tracking dyes that improve pipetting visibility and allow PCR products to be loaded directly onto an agarose gel without prior purification; amplified fragments have been successfully tested for cloning and restriction digestion without further cleanup.

Specifications include a concentration of 5 units/µl, recombinant enzyme status, extension rate of 2–4 kb/min at 72°C, half-life of 10 min at 97°C and 60 min at 94°C, amplification efficiency of ≥10^5-fold, 5'→3' exonuclease activity, extra A addition, and no 3'→5' exonuclease activity or contaminating nucleases, RNases, proteases, or self-priming activity. Substrate analogs accepted include dNTP, ddNTP, dUTP, biotin-11-dUTP, DIG-11-dUTP, and fluorescent-dNTP/ddNTP. The kit is suitable for PCR, RT-PCR, complex genomic templates, very low-copy targets, and genetic screening applications. Each lot is subjected to quality control tests including a stringent PCR specificity and reproducibility assay amplifying low-copy targets.