dnature diagnostics & research

Quantabio qScript® XLT One-Step RT-qPCR ToughMix®, 100 x 20 µl reactions

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qScript® XLT One-Step RT-qPCR ToughMix® 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® 5′-hydrolysis probes. First-strand cDNA synthesis and subsequent PCR amplification are carried out seamlessly in the same reaction mixture with optimized 1-step thermal cycling parameters. This kit 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 minimal reaction volumes and accelerated thermal cycling rates. The only necessary user-supplied materials for RT-qPCR is RNA sample and probe assay. It is compatible with all types of molecular probe assays including dual-labeling strategies. Elevating cDNA synthesis reaction temperature to 50-55°C during one-step RT-qPCR improves primer annealing and disruption of RNA secondary structure that can interfere with cDNA synthesis.

Features & Benefits

  • ToughMix reagent technology neutralizes a broad spectrum of PCR inhibitors
  • Superior assay sensitivity and specificity with ultrapure AccuStart II enzyme technology
  • Inert AccuVue plate loading dye simplifies reaction setup and provides instant visual cue of reagent addition and mixing
  • Supports efficient vortex mixing with proprietary anti-foaming formulation
  • Flexible – supports for both fast and standard thermal cycling conditions


2X reaction buffer containing

  • Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, dTTP
  • qScript XLT reverse transcriptase
  • RNase inhibitor protein
  • AccuStart II Taq DNA Polymerase
  • Inert AccuVue™ plate loading dye
  • Proprietary enzyme stabilizers and performance-enhancing additives
  • Titrated reference dye (if applicable)

Shipping & Storage

Store components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt.
Repeated freezing and thawing does not affect RT-qPCR performance.


Application notes for COVID-19 testing



Product Flyer

Application Note


E Gene Comparison between ToughMix

E Gene Comparison between ToughMix

E Gene comparing qScript 1-Step Virus ToughMix vs. qScript XLT 1-Step RT-qPCR ToughMix. 15, 150 and 1500 copies of RNA were compared. qScript 1-Step Virus ToughMix improves low copy RT-qPCR performance at both 150 and 15 copies of RNA.



  • I have been using qScript™ XLT One-Step RT-qPCR ToughMix for over 5 years- tens of thousands of reactions. Ease of use is an obvious major advantage, given it’s a straight single pipetting step to add it to the reaction mix, and it can be thawed on the bench and frozen back practically innumerable times. It also has comparatively short RT, and taq polymerase activation hold times. In six years or so of using this product we have come across many opportunities to verify other important features. Resilience: qScript™ XLT One-Step RT-qPCR ToughMix can be stored for 48 hrs. at 4⁰C without noticeable loss of activity- this finding was not by design. Versatility, in our hands, qScript™ XLT One-Step RT-qPCR Tough Mix works on NAs from a wide range of clinical veterinary samples, extracted using a variety of methods -including magnetic bead and spin column based systems. Samples include serum, blood, sputum and nasopharyngeal swabs, faeces (ruminant, canine, feline, avian) urine and various tissues (including fixed). We have used it as well for PCR only reactions, with identical performance to its PCR only ToughMix counterpart, which in turn out-performed other products we tested on DNA extracted from difficult (PCR inhibitory) samples. Practically, in our laboratory this means that we can duplex nucleic acid amplification of DNA and RNA. One of such Taqman duplex in our laboratory targets an RNA virus and a bacterium, with comparable performance to that of the individual reactions.
    We use half reactions (10 µL) which adds savings to the already very competitive pricing. Availability: always, and fast delivery. Very important for us as a pathology laboratory is that without exception all our urgent orders to dnature (numerous over 6 years) for this product have been delivered overnight. My experience with qScript™ XLT One-Step RT-qPCR ToughMix has been exceptionally good.
    GH – private laboratory

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