GoTaq® Probe 1-Step RT-qPCR System Technical Manual

Literature # TM379

The GoTaq® Probe 1-Step RT-qPCR System is optimized for quantitative PCR assays in the hydrolysis probe detection format. The system enables detection and relative quantification of RNA expression levels using a one-step RT-qPCR method, combining GoScript™ Reverse Transcriptase and GoTaq® Probe qPCR Master Mix in single-step real-time amplification reactions.

The GoScript™ RT Mix for 1-Step RT-qPCR (50X) combines optimized amounts of GoScript™ Reverse Transcriptase, RNasin® Plus RNase Inhibitor and additives to enhance single-step reactions. The GoTaq® Probe qPCR Master Mix included in the GoTaq® Probe 1-Step RT-qPCR System is formulated with dUTP. When dUTP is incorporated into the amplification products, the amplicons are susceptible to degradation by uracil-DNA glycosylase (UNG); this allows users to incorporate UNG into subsequent reactions for control of possible carryover contamination.

The GoTaq® Probe qPCR Master Mix is provided as a ready-to-use, stabilized 2X formulation that includes all components for qPCR (except template, primers and probe). This master mix does not contain a reference dye; however, a separate tube of carboxy-X-rhodamine (CXR) reference dye is included with this system, allowing users to add reference dye to amplification reactions if desired.