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Toy Block

OneStep RT-qPCR



  • Reverse transcription & qRT-PCR in one tube

  • Efficient cDNA synthesis ensured by the thermostable Reverse Transcriptase & advanced RNase Inhibitor mix

  • Works with high GC & high AT-rich templates

  • Rapid extension, early Ct detections

  • Very high sensitivity


  • Relative gene expression analysis

  • Absolute quantification of any RNA template (mRNA, total RNA, viral RNA), low copy number genes

  • qRT-PCR assays based on specific probes: including TaqMan®, Molecular Beacons, Scorpions™ Probes


  • 2X master mix for 500 reaction per 20µl reaction volume

  • Optimized with Hot Start Polymerase, Thermostable, and extremely active Reverse Transcriptase & advanced RNase Inhibitor allowing for a single step one tube qRT-PCR.

  • Highest sensitivity and specificity under both standard and fast qPCR cycling conditions.

  • Excellent performance on both AT/GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization.

  • Supplied with PCR Water to guaranty the best performance.

  • To cover all  RT-PCR instruments, the OneStep qRT-PCR Probe-based MasterMixes are available in three versions – without ROX, with low or high ROX concentration. See the ROX concentration table below to know which ROX concentration suits your instrument


Tips for successful workflow

  • Use special primer selection programs for good planning.

  • Work with amplicons in a range of 80-200, max 400 bp.

  • Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.

  • Run reactions in triplets; include a no-template control, no RT Mix control and positive control in parallel.

  • Thaw and keep reagents on ice. Mix well before use.

  • Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 °C temperature for this step.

To Prepare a 20 μl reaction, mix the following:

  • Reverse Primer: 100 - 400 nM final concentration.

  • Forward Primer: 100 - 400 nM final concentration.

  • Total RNA template: 1pg to 1 μg  (or mRNA template; >0.01 pg)

  • Add PCR Water to 10 μl volume

  • OneStep qRT-PCR MMix  (2X):  10 μl

  • RT3 Mix (20X)  1-2 μl  (2 μl improve Ct, but primer dimers may appear)

  • Mix gently, avoid bubbles. Place into the instrument

    • If Probe based MasterMix is used set the instrument channels according to the probes specifications

    • If Green MasterMix is used, set the instrument onto SYBR® Green or FAM channel

PCR Program

  • Reverse Transcription 1 cycle: 40-55°C - 10 min

  • Initial denaturation 1 cycle: 95°C - 2 min

  • Denaturation 40 cycles: 95°C - 5 sec

  • Annealing/extension 40 cycles: 60-65°C – 20-30 sec


Question regarding orders or bulk orders, please contact us

For technical questions about the products please contact our customer support

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