QuantiTect Probe RT-PCR Kit (1000)

A probe-based one-step RT-PCR master mix from QIAGEN designed for quantitative, real-time reverse-transcription PCR using sequence-specific probes. The kit enables accurate real-time quantification of RNA targets in an easy-to-handle format, combining reverse transcription and PCR in a single tube so that no tube opening is required once the reverse-transcription reaction has started.

The 2x QuantiTect Probe RT-PCR Master Mix contains HotStarTaq DNA Polymerase, a hot-start enzyme provided in an inactive state at ambient temperature, together with QuantiTect Probe RT-PCR Buffer (containing Tris·Cl, KCl, (NH4)2SO4, 8 mM MgCl2 at pH 8.7), a dNTP mix that includes dUTP, and ROX passive reference dye for fluorescence normalization on certain cyclers. The enzyme is activated by a 15-minute, 95°C incubation step after reverse transcription, which also inactivates the reverse-transcription enzymes and ensures temporal separation of the two steps in a single tube. The QuantiTect RT Mix provides an optimized blend of Omniscript Reverse Transcriptase (intended for RNA amounts greater than 50 ng) and Sensiscript Reverse Transcriptase (optimized for very small amounts of RNA, less than 50 ng). RNase-free water is also supplied.

The kit is suitable for use with all types of probe, including TaqMan dual-labeled probes, FRET probes, and Molecular Beacons, and has been optimized for real-time cyclers from Rotor-Gene, Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent. The buffer's balanced KCl and (NH4)2SO4 combination promotes stringent primer annealing, and Mg2+ titration is usually not required. The product is shipped on dry ice and should be stored at –20°C protected from light; the 2x master mix may alternatively be stored at 2–8°C for up to 6 months, while the QuantiTect RT Mix must always be kept at –20°C. The kit is intended for molecular biology applications and is not intended for the diagnosis, prevention, or treatment of disease.