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Quanta qScript™ XLT One-Step RT qPCR Toughmix, Low ROX 500 reactions

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95134-500

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Description

Protocol PDF

qScript XLT One-Step RT-qPCR ToughMix Low ROX is a ready-to-use, highly sensitive master mix for reverse transcription quantitative PCR (RT-qPCR) of RNA templates using hybridization probe detection chemistries such as TaqMan® hydrolysis probes on real-time PCR systems that require a passive reference dye. First-strand cDNA synthesis and PCR amplification are carried out in the same tube without opening between procedures.

It is ideal for highly sensitive quantification of RNA viruses or low abundance RNA targets as well as high throughput gene-expression studies. The system has been optimized to deliver maximum RT-PCR efficiency, sensitivity, and specificity in reduced reaction volumes and fast cycle times. qScript XLT One-Step RT-qPCR ToughMix ROX contains all required components for RT-qPCR except RNA template and probe. It is compatible with all dual-labeled probe chemistries.

qScript XLT is an engineered M-MLV reverse transcriptase with reduced RNase H activity and improved activity and stability at higher temperatures.

The use of higher temperatures (50 to 55°C) for the first-strand step of one-step RT-qPCR provides higher specificity for primer annealing and disruption of RNA secondary structure that can interfere with cDNA synthesis. qScript XLT One-Step RT-qPCR ToughMix Low ROX is highly resistant to PCR inhibitors. A key component of the ToughMix is an ultra pure, highly processive thermostable DNA polymerase that is combined with high avidity monoclonal antibodies. This provides an extremely stringent automatic hot-start that minimizes the potential for primer-dimer and other non-specific PCR artifacts. The light blue color of the AccuVue™ tracer dye simplifies reaction assembly in white, or clear, plates and helps to minimize pipetting or mixing errors. It does not interfere with qPCR performance or affect the stability of the product.

Features & Benefits

  • Tough – Overcomes common inhibitors including polysacharides, haeme/haemoglobin, humic acid & melanin
  • Flexible – Use fast or standard qPCR cycling conditions
  • Broad dynamic range – Reliable data from your precious samples every time
  • Multiplexing – supports detection of up to four targets


Real-Time PCR Instrument Compatibility

  • Applied Biosystems 7500
  • Applied Biosystems 7500 Fast
  • Stratagene Mx3000P®
  • Stratagene Mx3005P™
  • Stratagene Mx4000™
  • Applied Biosystems ViiA 7

Contents

qScript XLT One-Step RT-qPCR ToughMix Low ROX (2X):

2X reaction buffer containing dATP, dCTP, dGTP, dTTP, magnesium chloride, qScript XLT reverse transcriptase, RNase inhibitor protein, hot-start DNA polymerase, AccuVue blue qPCR dye, optimised ROX dye concentration and stabilisers

Shipping & Storage

qScript XLT One-Step RT-qPCR ToughMix Low ROX is stable for one year when stored in a constant temperature freezer at -20°C protected from light. Repeated freezing and thawing does not affect RT-qPCR performance.

 

PDF Protocol

 

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