Optimized Reagents for Probe-Based qPCR using the GoTaq® Probe qPCR and RT-qPCR Systems

21 Oct 2014

The GoTaq® Probe qPCR and RT-qPCR Systems are optimized for qPCR assays using probe-based detection chemistries. This article presents several experiments performed with the GoTaq® Probe qPCR Master Mix that exhibit sensitive quantification and robust performance.

GoScript™ Reverse Transcription System

Promega Corp.

The GoScript™ Reverse Transcription System includes a reverse transcriptase and a specialized set of reagents for efficient synthesis of first-strand cDNA optimized for quantitative PCR amplification. GoScript™ Reverse Transcriptase uses M-MLV Reverse Transcriptase and state-of-the-art buffer technology to deliver robust, reliable cDNA synthesis of a full range of rare and abundant transcripts, even in the presence of inhibitors. GoScript™ Reverse Transcriptase is qualified for use in qPCR, including GoTaq® qPCR and Plexor® RT-qPCR systems.GoScript™ Reverse Transcription System Features & Benefits: Ultra-Active - Save money on every reaction. Sensitive - Detect rare transcripts. Processive - Transcribe long messages. Resilient - Synthesize cDNA in the presence of strong inhibitors.

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GoTaq® Probe 1-Step RT-qPCR System

Promega Corp.

  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 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. The GoTaq® Probe qPCR Master Mix is designed to provide resistance to a wide range of PCR inhibitors. This formulation uses antibody-mediated hot-start chemistry, allowing reaction setup to be performed at room temperature. The master mix also employs rapid hot-start activation and processive enzymes, making it compatible with both standard and fast instrument cycling programs. Features - Benefits • Superior Performance: Sensitive detection on any real-time instrument • Enhanced Stability: Exceptional room-temperature setup makes it suitable for automation and high-throughput detection • Versatility: Compatible with both fast and standard cycling methods • Robustness: High-efficiency, full-length cDNA synthesis in the presence of inhibitors • Confidence: The Promega PCR Performance Guarantee Applications • Sensitive sequence detection and quantification • Gene expression analysis • Detection of sequence variants • Fluorescent detection  

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GoTaq® Probe qPCR Master Mix

Promega Corp.

The GoTaq® Probe qPCR Master Mix is optimized for quantitative PCR assays in the hydrolysis probe detection format.   It 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. The GoTaq® Probe qPCR Master Mix is designed to provide resistance to a wide range of PCR inhibitors. This formulation uses antibody-mediated hot-start chemistry, allowing reaction setup to be performed at room temperature. The master mix also employs rapid hot-start activation and processive enzymes, making it compatible with both standard and fast instrument cycling programs. Features - Benefits • Superior Performance: Sensitive detection on any real-time instrument • Enhanced Stability: Exceptional room-temperature setup makes it suitable for automation and high-throughput detection • Versatility: Compatible with both fast and standard cycling methods • Robustness: High-efficiency, full-length cDNA synthesis in the presence of inhibitors • Confidence: The Promega PCR Performance Guarantee Applications • Sensitive sequence detection and quantification • Gene expression analysis • Detection of sequence variants • Fluorescent detection  

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