qPCR Master Mix Kit, 1mL

For laboratories running TaqMan probe-based quantitative PCR, this 2X-concentrated mix from STEMCELL Technologies combines a hot-start DNA polymerase, dNTPs, MgCl2, enhancers, and stabilizers in a single vial for cDNA amplification. Each 1 mL kit ships with a separate 200 µL vial of ROX Reference Dye (companion catalog #07519), giving users the option to configure the assay for ROX-dependent and ROX-independent real-time instruments. The protocol recommends 5 µL of master mix per 10 µL reaction with 250 to 1000 nM forward and reverse primers and 150 to 250 nM probe, and accommodates a DNA template input range of 3 pg to 100 ng per well.

Researchers routinely apply this reagent to gene expression analysis, mRNA quantification, validation of RNA-seq results, genotyping, and pathway studies under standard or fast cycling conditions. Because the hot-start enzyme remains inactive at room temperature, reaction plates can be assembled at the bench before thermal cycling. After loading, centrifuge the sealed plate at 1000 x g for 1 minute at room temperature (15 to 25°C) to dislodge bubbles from the well bottoms before transferring to the block.

Both components ship frozen and should be stored at -20°C, protected from light, and remain stable until the expiry date printed on the label. Thaw on ice and invert gently to mix without foaming; the master mix tolerates repeated freeze-thaw cycles. The mix is compatible with real-time thermal cyclers from multiple manufacturers and is intended for research use only.