dnature diagnostics & research

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

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95132-500

SKU: 95132-500 Categories: , ,

Description

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® hydrolysis probes on real-time PCR systems that do not 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 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 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

  • Roche LightCycler™ 480
  • Qiagen/Corbett Rotor-Gene®
  • Illumina Eco
  • MIC
  • Eppendorf Mastercycler® ep realplex
  • Cepheid SmartCycler®
  • Bio-Rad CFX96™
  • Bio-Rad CFX384™

Contents

qScript XLT One-Step RT-qPCR ToughMix (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, and stabilisers

Shipping & Storage

qScript XLT One-Step RT-qPCR ToughMix 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

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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