Reverse transcription quantitative PCR (RT-qPCR) is the method of choice for accurate, sensitive, and fast gene expression analysis. With more and more researchers relying on PrimeTime qPCR Assays and Probes, we recognized the need for a quality gene expression master mix to go with.
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).