• Fast: Complete cDNA synthesis and robust amplification in as little as 1.5 hours
  • Flexible: Compatible with both SYBR-Green and TaqMan Probe-based assays
  • Efficient: Superior performance compared to competitor RT-qPCR kits

ZymoScript One-Step RT-qPCR Kit contains all the necessary components for fast reverse transcription and robust qPCR amplification in a single kit. Room-temperature reaction setup is quick and easy. The ZymoScript One-Step RT-qPCR Kit eliminates the need to transfer tubes between reverse transcription and qPCR, which minimizes sample handling and reduces the risk of contamination.

The included Reaction Buffer (4X) contains dNTPs, MgCl2, and other additives to ensure complete cDNA synthesis and specific qPCR amplification in just 1.5 hours. Non-interfering blue tracking dye is included to simplify reaction setup and prevent pipetting errors. The 20X Enzyme Mix containing temperature-activated reverse transcriptase, hot-start DNA polymerase, and RNase inhibitors ensures maximal specificity. The ZymoScript One-Step RT-qPCR kit is compatible with both SYBR-Green and TaqMan probe-based assays for ultimate flexibility. A 20X Fluor Dye for SYBR-Green based detection methods is included for added convenience. Detection of multiple targets in a single reaction (multiplex RT-qPCR) is feasible using TaqMan probes (not provided).

The ZymoScript One-Step RT-qPCR Kit has been rigorously tested to demonstrate superior sensitivity, specificity, and reproducibility. The kit can reliably quantify as little as 0.1 pg RNA (including difficult GC-rich targets) and outperforms other commercial kits. This kit was also used to develop a molecular diagnostic test for COVID-19 ranked third in sensitivity by the American FDA. The unique buffer system and temperature-activated enzymes facilitate the melting of secondary structures and ensure ultimate specificity for the intended product. ZymoScript One-Step RT-qPCR Kit is the product of choice for simple and sensitive RT-qPCR. It is the ideal product to use as a building block for RT-qPCR multiplex assays or high-throughput diagnostic applications. For quick and efficient cDNA synthesis and 2-step RT-qPCR options, see ZymoScript RT PreMix Kit (R3012) and ZymoTaq qPCR Premix (E2054).

Technical Specifications
Amplicon Length This product performs best with amplicons between 50 bp and 1.5 kb
Equipment thermal cycler/qPCR instrument
Ideal Uses high-throughput applications, multiplex diagnostic assays based on RNA targets, routine quantification of specific targets in multiple samples
Processing Time as little as 1.5 hours
Reagents Complete and ready-to-use Reaction Buffer (4X) and Enzyme Mix (20X). Fluor Dye (20X) included for SYBR-Green based detection. MgCl2 included to supplement the magnesium concentration if needed.
Sample Source 0.1 pg – 5µg RNA input
Size 100 reactions
Storage Store at -20 °C. Minimize exposure to light.
Supplemental Info


The amount of total RNA required may vary depending on the expression level of the target transcripts. In general, we recommend using 0.1 pg – 5 µg of input RNA.

The dye used in the ZymoScript One-Step RT-qPCR Kit has been thoroughly tested in downstream analysis. At the provided concentration, it will not affect fluorescent signal readings during qPCR.

Lower signal can be expected with degraded and impure RNA samples. We recommend using Quick-RNA or Direct-zol Kits (see Related Products) for high quality RNA extraction.

No, only use gene specific primers when using ZymoScript One-Step RT-qPCR Kit.

Yes, the reaction can be assembled at room temperature. The enzymes are only activated at higher temperatures, ensuring maximum specificity.

We do not recommend storing the kit or kit components at room temperature for prolonged periods of time. The kit can be stored at 4°C for up to 1 week.

Samples can be treated with DNase I to eliminate DNA contamination. DNase I is included in the Quick-RNA or Direct-zol Kits recommended for RNA extraction. Alternatively, the DNase I set (see Related Products) can be purchased separately.

No, RNase inhibitors are already included in the enzyme mix to prevent unwanted RNA degradation during reverse transcription.

This depends on different factors, including RNA integrity and primer efficiencies. Generally, this product performs the best with amplicons with a length between 50 bp and 1.5 Kb.

product video

ZymoScript One-Step RT-qPCR Kit

Cat # Name Size
R3014 ZymoScript One-Step RT-qPCR Kit 100 reactions