New Two-Step PCR System Improves Quantification Sensitivity

A new two-step kit has been placed on the market for biotech researchers wishing to detect the activity of target genes expressed at low levels, even with hard-to-amplify samples.

The new Promega (Madison, WI, USA) GoTaq 2-Step RT-qPCR system comprises two Promega test modules: the GoScript Reverse Transcription System and the GoTaq qPCR Master Mix.

In the first step, by capitalizing on a carefully blended set of reagents the GoScript Reverse Transcriptase module directs highly efficient full-length complementary DNA (cDNA) synthesis from rare and abundant targets with up to five micrograms of RNA input. The transcription system improves the yield of cDNA even in the presence of common inhibitors such as ethanol.

In the second step, cDNA is quantified using the GoTaq qPCR Master Mix. Formulated with a hot-start DNA polymerase, the GoTaq qPCR Master Mix is optimized for reproducibly specific, sensitive, highly efficient qPCR using the standard or fast mode of a real-time PCR instrument. The GoTaq qPCR Master Mix module contains the green dye BRTY Green, which can fluoresce at twice brightness of the commonly used SYBR Green I dye.

The GoTaq 2-Step RT-qPCR System is a ready-to-use kit for analyzing a wide range of RNA targets by combining the high-activity of GoScript Reverse Transcription System with the ultrabright fluorescence of GoTaq qPCR Master Mix. The system is robust, requires little optimization, and is flexible for a variety of applications.

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