Essentials of Real-time PCR
A general overview of the basics of real-time PCR (qPCR) assays, sequence detection chemistries, TaqMan and SYBR chemistries, quantitation assays, and methods of absolute vs. relative quantitation.
Whether you're new to real-time PCR, also called quantitative PCR (qPCR), or want to learn about new applications for real-time PCR, we have the learning material, including videos and webinars, to help you understand the technology and get started quickly.
Real-time PCR combines PCR amplification and detection into a single step. This eliminates the need to detect products using gel electrophoresis, and more importantly it enables the method to be truly quantitative. With real-time PCR, fluorescent dyes are used to label PCR products during thermal cycling. Real-time PCR instruments measure the accumulation of fluorescent signal during the exponential phase of the reaction for fast, precise quantification of PCR products and objective data analysis.
Featured resources
Handbook
Real-Time PCR Handbook 2.0
An excellent resource for anyone who is new to real-time PCR and interested in learning the basic principles of the technology. Our newly released edition for 2020 provides a rich digital experience with embedded videos and even more resources to help you dig deeper into topics of interest such as qPCR basics, assay design, data analysis, troubleshooting, and more.
Poster
Real-time PCR Taq Talk video series poster
A free interactive poster that provides a simple and intuitive pathway to a robust library of qPCR educational videos and guidance through your research workflow.
Videos
Taq Talk video series
As experts in real-time PCR, the Taq Team will address your most frequently asked real-time PCR questions and challenges in a new video series, Taq Talk, where viewers can expect to hear everything from the basics and best practices to experiment setup and choosing the right master mix.
Taq FAQ: How to load an Applied Biosystems™ TaqMan™ Array Card
Watch this short video to see just how easy it is to load a TaqMan Array Card, featuring Nathan Zuzow, PhD, Field Application Scientist at Thermo Fisher Scientific. Stay tuned for more tips from the field.
Video: What can you study using real-time PCR?
Real-time PCR, also known as qPCR, is used for many qualitative and quantitative applications, including gene expression analysis, microRNA analysis for identification of cancer biomarkers, single nucleotide polymorphism (SNP) genotyping, copy number variation (CNV) analysis, and even protein analysis. Taq Talk Episode 15 covers these common real-time PCR applications.
What is UNG/UDG?
UNG or UDG are used in many master mixes. This article describes the two enzymes and how they are used in qPCR.
Good Laboratory Practice to Avoid Contamination
Learn how to spot contamination issues and to avoid or reduce contamination risks
Basic Principles of RT-qPCR
When choosing a reverse transcription method, you want to ask yourself several questions, Do I want to do 1-step or 2-step? Do I want to start with total RNA or mRNA? And finally, what type of RT primer do I want to use?
Real-Time PCR: Understanding Ct
This review will highlight the factors that must be considered when setting up and evaluating a real-time PCR (qPCR) reaction.
Absolute vs. Relative Quantification for Real-Time PCR
In absolute quantification using digital PCR, no known standards are needed. The target of interest can be directly quantified with precision determined by number of digital PCR replicates. In relative quantification, you analyze changes in gene expression in a given sample relative to another reference sample (such as an untreated control sample).
How TaqMan Assays Work
A brief explanation and illustration of how the TaqMan chemistry works in the various applications of gene expression, genotyping, copy number variation, microRNA, mutation detection, and protein quantification.
TaqMan Assays Help Meet Your Research Needs
Depending on your research needs, you can find the best TaqMan Assay and format to enable best-in-class performance for applications requiring the utmost specificity and sensitivity.
Article
Calculating Primer and Probe Concentrations
Follow example calculations to prepare and use oligonucleotide working stocks with confidence and precision.
Article
Common TaqMan Myths Busted
How much do you know about TaqMan Assays? Discover some of the common misconceptions researchers have about this chemistry.
Additional resources
- Real-time PCR Taq Talk video series poster
- Blog: TaqMan Assays – 5 Interesting Facts
- Blog: A Quick Guide to qPCR Terms and Acronyms
- Real-Time PCR webinars
- Genetic Analysis for All webinars
- TaqMan Gene Expression Assays brochure
- TaqMan Assays infographic
- Using TaqMan Endogenous Control Assays to Select an Endogenous Control for Experimental Studies
- The design process of quantitative TaqMan gene expression analysis tools
- Real-Time vs. Digital vs. Traditional PCR
- TaqMan vs. SYBR Chemistry for Real-Time PCR
- Why TaqMan Assay Design is Important
- Efficiency of Real-Time PCR (qPCR)
- What is multiplex qPCR?
Real-Time PCR Resource Library
Innovative tools and educational resources that help advance the work of researchers engaged in real-time PCR.
For Research Use Only. Not for use in diagnostic procedures.