QuantiTect Multiplex PCR Kit (200)

From QIAGEN, this kit enables quantitative, multiplex, real-time PCR and two-step RT-PCR using sequence-specific probes. The 2x QuantiTect Multiplex PCR Master Mix contains HotStarTaq DNA Polymerase, QuantiTect Multiplex PCR Buffer with 11 mM MgCl2, a dNTP mix of dATP, dCTP, dGTP, and dTTP/dUTP, and ROX passive reference dye. The kit is optimized for use with real-time cyclers that require ROX for fluorescence normalization, including instruments from Applied Biosystems.

The hot-start enzyme HotStarTaq DNA Polymerase is a modified form of a recombinant 94 kDa DNA polymerase originally isolated from Thermus aquaticus. It is provided inactive at ambient temperature, preventing the formation of misprimed products and primer–dimers during reaction setup, and is activated by a 15-minute, 95°C incubation step. The kit supports duplex, triplex or 4-plex PCR, with up to 5 targets quantifiable simultaneously depending on the cycler. Its inclusion of dUTP allows optional uracil-N-glycosylase (UNG) pretreatment where carry-over contamination is a concern.

The specialized buffer delivers high specificity and sensitivity without time-consuming optimization, and the kit is compatible with all types of probe chemistries. The kit should be stored at –30 to –15°C (per the quick-start protocol) or –20°C (per the handbook), protected from light; the master mix may alternatively be stored at 2–8°C for up to 6 months.