PCR and qPCR
Support and Educational Content

Improve probe-based qPCR experiments

PrimeTime® Gene Expression Master Mix



Performance data and guidance for better qPCR research

Real-time quantitative PCR (qPCR) is a mainstream method for accurate, sensitive, and fast gene expression analysis that is used in research and diagnostic applications. PrimeTime Gene Expression Master Mix for 5′ nuclease assays has been optimized for use with PrimeTime qPCR Assays—it also performs well with probe-based assays from other manufacturers. This versatile master mix is compatible with a wide range of instruments, with or without a reference dye, and can be used under standard or fast cycling conditions. PrimeTime master mix is extremely stable, delivering consistent qPCR performance in singleplex and multiplex reactions in overnight, high throughput experiments.

In the webinar, High efficiency qPCR with the PrimeTime® Gene Expression Master Mix from IDT, Dr Erik Wendlandt presents performance data, discusses key considerations for singleplex and multiplex experimental formats, and describes online tools that can help you plan your qPCR experiments. Access the recorded presentation below to learn about performing successful gene expression experiments with PrimeTime Gene Expression Master Mix. At the end of the recording, Dr Wendlandt answers questions received from webinar attendees.

Product focus

PrimeTime® Gene Expression Master Mix

For use with probe-based qPCR assays.

  • Achieve high efficiency qPCR under fast or standard cycling conditions
  • Multiplex without loss of sensitivity
  • Obtain consistent results by capitalizing on exceptional benchtop stability

Learn more at www.idtdna.com/qPCRMasterMix. For information about license-free options for commercial or diagnostic use, contact custcare@idtdna.com.


PrimeTime qPCR Assays

  • 5′ nuclease, probe-based assays—the gold standard for quantitative gene expression studies
  • Primer-based assays—designed for intercalating dye experiments (Note: currently, not for use with PrimeTime Gene Expression Master Mix)

Design custom assays for any target to your specific parameters, using our proprietary bioinformatics algorithms. Alternatively, select from our predesigned assays for human, mouse, and rat mRNA targets that are supported by our bioinformatics algorithms and up-to-date sequence information.

Learn more at www.idtdna.com/PrimeTime. For assistance with assay design, contact our scientific application specialists at applicationsupport@idtdna.com.


Double-Quenched Probes

ZEN™ and TAO™ Double-Quenched Probes have a 5′ fluorophore, an internal quencher (ZEN or TAO quencher), and a 3′ quencher (Iowa Black® FQ). These probes provide consistently earlier Cq values and improved precision, when compared to traditional, single-quenched qPCR probes.

Learn more at www.idtdna.com/qPCRprobes.


gBlocks® Gene Fragments

gBlocks Gene Fragments are double-stranded, 125–2000 bp DNA molecules. They are ideal for use as qPCR controls and standards, as well as for gene construction and editing applications. These affordable gene fragments are sequence-verified and ship in a few working days, saving valuable laboratory time.

Learn more at www.idtdna.com/gBlocks.

Further reading

Design efficient PCR and qPCR primers and probes using online tools—Find out how to facilitate experimental planning using IDT free, online tools for oligonucleotide analysis and PCR primer design. This article describes our predesigned qPCR assays and the basics of designing customized PCR primers and hydrolysis probes with the PrimerQuest® Tool.


Increase sensitivity and precision in your qPCR experiments—Use double-quenched probes to decrease background, and increase sensitivity and precision in your qPCR experiments.


Easily-designed standard curves for qPCR—Learn how to use synthetic gBlocks Gene Fragments for creating standard curves, including incorporating multiple targets into a single gene fragment.


Multiplex qPCR—how to get started—Review tips for setting up multiplex qPCR experiments.


Recommended dye combinations for multiplex qPCR—Read these recommendations for selecting dyes for multiplex qPCR to minimize background and avoid overlap of fluorescent signals. Included is a table of compatible dyes for multiplexing on common qPCR instruments and a list of suggested quenchers.

Author: Hans Packer, PhD, is a scientific writer at IDT.

© 2016 Integrated DNA Technologies. All rights reserved. Trademarks contained herein are the property of Integrated DNA Technologies, Inc. or their respective owners. For specific trademark and licensing information, see www.idtdna.com/trademarks.


Predesigned qPCR Assays

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

Search human, mouse, or rat genes ≫


Related Articles

Designing PCR Primers and Probes

General guidelines for designing primers and probes and for choosing target locations for PCR amplification.

Read more ≫

Steps for a Successful qPCR Experiment

Considerations for 5′ nuclease assay design and experimental setup to help you obtain accurate and consistent results.

Read more ≫

Interpreting Melt Curves: An Indicator, Not a Diagnosis

Examining PCR melt curve data to determine what it can/cannot tell us about resulting PCR amplicons.

Read more ≫

Epigenetic Biomarkers for Prostate Cancer

Scientists use methylation and expression analysis methods to evaluate epigenetic markers for early, noninvasive detection of aggressive prostate cancer. IDT PrimeTime® qPCR Assays, ZEN™ Double-Quenched Probes, and gBlocks® Gene Fragments facilitate this research.

Read more ≫

Optimizing Multiplex qPCR for Detecting Infectious Diseases and Biothreat Agents in the Field—ZEN™ Double-Quenched Probes bring down the background

Tetracore researchers developing large sets of robust probe-based qPCR assays discuss the need to: use probe dyes compatible on common PCR instruments, maintain low background with multiple probes, and reformulate assays to address viral mutation.

Read more ≫