QuantiTect Probe PCR Kit (200)

QIAGEN's probe-based real-time PCR kit enables sensitive quantification of genomic DNA and cDNA targets using dual-labeled sequence-specific probes, including TaqMan® probes, FRET probes, and Molecular Beacons. The kit provides a ready-to-use 2x QuantiTect Probe PCR Master Mix that contains HotStarTaq® DNA Polymerase, activated by an initial 15-minute incubation at 95 °C to deliver a stringent hot start, and a proprietary PCR buffer with a balanced combination of K⁺ and NH₄⁺ ions that promotes specific primer annealing. The master mix also includes ROX passive reference dye for fluorescence normalization on Applied Biosystems® cyclers and optionally on Agilent instruments, and a dNTP mix containing dUTP, which allows optional uracil-N-glycosylase (UNG) pretreatment to eliminate carry-over contamination from previous PCR products.

The kit is compatible with most real-time thermal cyclers, including the LightCycler®, Rotor-Gene®, Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent/Stratagene instruments. The protocol recommends an initial Mg²⁺ concentration of 4 mM as provided by the master mix, and for highest efficiency targets should ideally be 100–150 bp in length. Because the hot-start formulation eliminates nonspecific amplification at room temperature, reactions can be set up at the bench without keeping samples on ice. The product is manufactured under Good Manufacturing Practice (GMP) principles to ensure consistent performance. On receipt, the kit should be stored at –30 to –15 °C in a constant-temperature freezer and protected from light; the 2x QuantiTect Probe PCR Master Mix may also be stored protected from light at 2–8 °C for up to 6 months, depending on the expiration date.