ZymoScript One-Step RT-qPCR Kit
R3014
ZymoScript One-Step RT-qPCR Kit
Cat # | Name | Size | Price | Quantity |
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Documents

Highlights
- 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
Description
Amplicon Length | This product performs best with amplicons between 50 bp and 1.5 kb |
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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 |
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Q1: What is the minimum/maximum amount of RNA template I should use?
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.
Q2: Will the blue tracking dye interfere with qPCR fluorophore channels and affect the signal of probe/fluorescent dyes?
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.
Q3: How is the performance of the ZymoScript One-Step RT-qPCR kit with low-quality RNA samples?
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.
Q4: RT-qPCR signal is not detected, or it is detected at higher Ct than expected. What should I do?
A possible reason for missing or delayed amplification is inefficient primer binding to the template. Make sure to use an annealing temperature (step 4 in the protocol provided) at least 2°C below the lowest melting temperature of the primers to ensure efficient binding to the target sequence.
Q5: Can I assemble the RT reaction at room temperature?
Yes, the reaction can be assembled at room temperature. The enzymes are only activated at higher temperatures, ensuring maximum specificity.
Q6: Can I store the ZymoScript One-Step RT-qPCR kit at Room Temperature or 4°C?
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.
Q7: How can I ensure that there is no genomic DNA contamination in my RT reaction?
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.
Q8: Do I need to add RNase inhibitors?
No, RNase inhibitors are already included in the enzyme mix to prevent unwanted RNA degradation during reverse transcription.
Q9: Should I use SYBR Green or a TaqMan probe-based assay?
For single target detection, SYBR Green based assays offer a more affordable solution. For SYBR Green based assays, add 1 µl of Fluor Dye solution (20X) to each 20 µl RT-qPCR reaction. TaqMan probe-based assays are in general more expensive than SYBR Green based assays. However, they present an advantage when multiple targets are analyzed simultaneously and when a superior level of specificity is required. Do not add Fluor Dye solution to the RT-qPCR reaction when performing TaqMan probe-based assays.
Q10: Which channel should be activated when using the Fluor Dye solution?
The Fluor Dye has the same excitation and emission properties as SYBR Green. Activate the SYBR/FAM channel for detection when using the Fluor Dye.
Q11: Does ZymoScript One-Step RT-qPCR contain a passive reference dye?
No, a passive reference dye is not included.
Q12: What is the amplicon length that can be achieved using the ZymoScript One-Step RT-qPCR Kit?
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.