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 ZymoScript RT PreMix has been thoroughly tested in downstream analyses. At the provided concentration, it will not affect fluorescent signal readings of any qPCR fluorescent channel.

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

Yes, gene specific primers can be used to increase specific target yield. However, we recommend using ZymoScript One Step RT-qPCR (see Related Products) for enhanced specificity.

Yes, the enzymes are not activated at ambient temperatures.

We do not recommend storing the kit at room temperature for prolonged periods of time. The RT PreMix 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 and Direct-zol Kits recommended for RNA extraction (see Related Products). Alternatively, the DNase I set can be purchased separately.

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

No. The ZymoScript RT PreMix (2X) already contains a mixture of oligo dT and random hexamer primers which is not ideal for the synthesis of long cDNA molecules. For this specific application we recommend using the ZymoScript One-Step RT-qPCR Kit (R3014) with gene-specific primers.