QuantiTect Probe PCR Kit (1000x50µl)

Designed for real-time quantification of gDNA and cDNA targets using dual-labeled probes such as TaqMan, FRET, and Molecular Beacon probes, this PCR kit from QIAGEN is manufactured under Good Manufacturing Practice (GMP) principles to ensure consistent product performance.

The 2x QuantiTect Probe PCR Master Mix supplied in the kit contains ROX passive reference dye — required for all real-time cyclers from Applied Biosystems but compatible with all real-time instruments, as it does not interfere with PCR on any other cycler — as well as dUTP, which permits optional uracil-N-glycosylase (UNG) pretreatment of the reaction. The mix also provides HotStarTaq DNA Polymerase, which requires an initial 15 min incubation at 95°C to activate. The kit is suitable for use with most real-time cyclers, including the LightCycler 1.x and 2.0, Rotor-Gene, and instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent/Stratagene.

For optimal amplification efficiency, target sequences should ideally be 100–150 bp in length. The protocol recommends starting with the 4 mM Mg2+ concentration provided by the master mix, though concentrations up to 6 mM may be tested for a very limited number of targets. Standard cycling uses an optional 2 min UNG step at 50°C, followed by 35–55 cycles on LightCycler instruments or 35–45 cycles on other cyclers, with denaturation at 95°C (LightCycler) or 94°C (other cyclers) and a combined annealing/extension step at 60°C for 60 s. The kit should be stored at –30 to –15°C, protected from light; the master mix may also be kept at 2–8°C for up to 6 months, depending on the expiration date.