dnature diagnostics & research

Quantabio qScript® 1-Step Virus ToughMix®, 500 x 20 µl reactions

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95131-500

Description

qScript® 1-Step Virus ToughMix® is a 2x, ready-to-use, master mix for rapid detection of RNA viruses such as Flu-A, Flu-B, and SARS-CoV-2, using one-step, or single-tube reverse transcription quantitative PCR (RT-qPCR). It has been optimized for maximum sensitivity to enable reliable quantification of very low input quantities of RNA using dual-labeled hydrolysis probe detection chemistries such as TaqMan® probes in single or multiplexed assay formats. qScript® 1-Step Virus ToughMix® contains all required components for RT-qPCR except RNA template and primer/probe assays.

qScript® 1-Step Virus ToughMix® is powered by an engineered reverse transcriptase with reduced RNase H activity and improved activity and stability at higher temperatures, that includes a RT Hot-Start technology to suppresses non-specific primer extension by the RT before cDNA synthesis. 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. These features combined with a stringent mAb hot-start Taq polymerase and ToughMix reagent technology to neutralize many common PCR inhibitors, provide reproducible low-copy quantification as well as extended room temperature stability of fully assembled reactions for automated reaction assembly.

qScript® 1-Step Virus ToughMix® does not contain an internal reference dye.

Features & Benefits

  • Superior Sensitivity – higher amplification yields enable lower copy detection
  • Tough Tested – tolerant to a wide range of PCR inhibitors
  • Multiplexing – highly sensitive detection for up to four targets

Components

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, and stabilizers

Shipping & Storage

Store components in a constant temperature freezer at -25°C to -15°C. Repeated freezing and thawing do not affect RT-qPCR performance.

Application notes for COVID-19 testing

Protocol

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.

 

Reviews

  • 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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