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qPCR with intercalating dyes: PrimeTime® qPCR Primers

PrimeTime® qPCR Primers are primer sets predesigned to detect genes in human, mouse, and rat transcriptomes. These primer sets are ideal for use with intercalating dyes such as SYBR® Green and EvaGreen®, where no probe is needed, and they eliminate the inconvenience of designing specific assays for intercalating dyes. Now you can simplify the transition from discovery to screening by using PrimeTime qPCR Primers to analyze large sample numbers in initial discovery before upgrading to probe-based assays (PrimeTime qPCR Assays) for improved specificity and performance as sample numbers decrease.

The benefits of PrimeTime qPCR Primers include:

  • Up-to-date sequence design—While primer binding sites are carefully pre-chosen using a sophisticated design engine, primers are only synthesized at the time of order using current sequence information. A BLAST search is also performed to eliminate off-target effects.
  • Guaranteed performance, high efficiency—Average qPCR efficiencies are >90% and gel analysis consistently shows single bands for PCR products.
  • Compatible with popular master mixes—Comparable efficiencies are achieved when the primers are used with various commercial master mixes under manufacturers’ cycling conditions. See the data under PrimeTime Product Performance.
  • Simple ordering and sequence provided—Simple assay selection interface with sequence information provided upon order placement.
  • Easy transition to probe-based assays—PrimeTime qPCR Primers use the same designs as IDT PrimeTime probe-based Assays. So when you are ready to move to 5' nuclease assays, you can just request the associated probe for those same primer sequences.

To order, use the PrimeTime Predesigned qPCR Assay Database or the RealTime PCR Tool. You can also use the RealTime PCR Tool to design and order custom primers (Note: The performance of custom designed primers is not guaranteed by IDT). PrimeTime qPCR Primers ship in 2–3 business days.

Product focus: qPCR Reagents—everything but your sample

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

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Interpreting melt curves: An indicator, not a diagnosis—Performing intercalating dye PCR/qPCR assays? Review examples of PCR melt curve data with our scientists to determine what it can/cannot tell us about resulting PCR amplicons.

Melt curve analysis for improved intercalating dye qPCR—Listen to this discussion of the benefits and limitations of melt-curve analysis using clear example—presented as a webinar. Read how PrimeTime Predesigned qPCR Primer Assays are designed to be compatible with intercalating dyes, and can also be transitioned to probe-based assays.

Tips for using BLAST to locate PCR primers—Need a quick way to find the location of primers within a gene or the expected size of the resultant PCR product? In this tip we show you how to get this information using BLAST.

SYBR® to 5' nuclease assays—Product spotlight: Researchers often use intercalating dyes, like SYBR® Green I, for qPCR experiments for the advantage they bring in lowered price and reduced turnaround time. However, the disadvantages to using these reagents are significant. Read how easy it is to convert to 5’ nuclease assays and the advantages the addition of a hydrolysis probe will provide.

Author: Nicola Brookman-Amissah is a senior scientific writer at IDT.

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