Real-Time PCR Basics | Thermo Fisher Scientific

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

Optimized protocol with low sample input for profiling human microRNA

Real-Time PCR Handbook

An excellent resource for anyone who is new to real-time PCR and interested in learning the basic principles of the technology. This 3rd edition contains 6 chapters in 70 pages and also covers topics such as assay design, data analysis, real-time PCR tips, and troubleshooting.

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.

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.

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’s the difference between qPCR vs. Digital PCR vs. Traditional PCR?

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.

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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).