PCR and qPCR
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Obtain high efficiency qPCR results using PrimeTime® Gene Expression Master Mix

For probe-based qPCR assays

Reverse transcription quantitative PCR (RT-qPCR) is the method of choice for accurate, sensitive, and fast gene expression analysis. Many of you have come to rely on IDT for efficient PrimeTime qPCR Assays and Probes. Now you can consistently achieve high efficiency results using our gene expression master mix in your qPCRs.

PrimeTime Gene Expression Master Mix is a versatile and convenient master mix for 5′ nuclease assays. It has been optimized for qPCR and 2-step RT-qPCR using PrimeTime qPCR assays, but also performs well with primer–probe sets from IDT and other manufacturers. In addition, this master mix is compatible with a wide range of instruments—whether you prefer standard or fast cycling conditions, and whether or not you need a reference dye.

Reproducible results for high-throughput or overnight applications, every time

PrimeTime Gene Expression Master Mix has been formulated for use at room temperature, so you can let your reactions cycle overnight or perform automated, high-throughput experiments (Figure 1). Cq values were consistent, when qPCRs were run immediately or after 24 hr at room temperature. The standard deviations for the Cq values were <0.5 for template levels >10 copies. This result was obtained with 3 different lots of master mix, so you can rely on this reagent to provide reproducible results with qPCR assays.

Figure 1. Consistent results at 0 and 24 hours across different lots of PrimeTime® Gene Expression Master Mix. qPCR mixes of PrimeTime HPRT Assay (Assay ID: Hs.PT.58v.45621572), PrimeTime Gene Expression Master Mix (from 1 of 3 lots), reference dye, and varying amounts of gBlocks® Gene Fragments (10–107 copies; 8 replicates) were assembled. A JANUS® automated, liquid-handling workstation (PerkinElmer) was used to load the reactions into PCR plates. Reactions were run immediately (Day 0) or after 24 hours (Day 1) on a QuantStudio™ 7 Flex System (Thermo Fisher Scientific).

Learn more about PrimeTime® Gene Expression Master Mix.

PrimeTime Gene Expression Master Mix is already advancing agricultural testing:

"The PrimeTime Gene Expression Master Mix worked very well for us. I was able to use it effectively in a 2-target multiplex reaction, with no difference in amplification of the targets compared to the singleplex reactions."

 – Sandy Junek, Molecular Lab Manager, Discovery Seed Labs (Saskatoon, SK, Canada)

Product focus: Everything but your sample

All the reagents you need for successful qPCR assays are now available through IDT.

Additional resources

Application guide: PrimeTime qPCR Application Guide, 4th Ed.—Download this useful resource that provides experimental overviews, protocols, data analysis, and troubleshooting chapters.

Article: Design efficient PCR and qPCR primers and probes using online tools—To facilitate experimental planning, IDT offers a variety of free, online tools for oligonucleotide analysis and PCR primer design. This article provides an overview of our predesigned qPCR assays and the basics of designing customized PCR primers and hydrolysis probes with the PrimerQuest® Tool.

Webinar: Strategies for optimizing high throughput qPCR for expression profiling—Learn how to address the challenges of high throughput RT-qPCR expression profiling from a prominent qPCR expert, Dr Mikael Kubista (TATAA Biocenter, Sweden).

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Predesigned qPCR Assays

Probe-based qPCR assays for quantification of human, mouse, and rat gene expression. Order in plates or tubes.

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