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.
Real-Time PCR (qPCR) Basics
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.
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.
Get answers to popular qPCR questions with our library of quick and easy how-to videos covering a variety of topics including experiment setup, best practices, and guidance on products and technologies.
Video: What can you do with real-time PCR (qPCR) applications?
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. This Ask TaqMan Ep. 17 video covers these common real-time PCR applications.
UNG or UDG are used in many master mixes. This article describes the two enzymes and how they are used in qPCR.
Learn how to spot contamination issues and to avoid or reduce contamination risks
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?
This review will highlight the factors that must be considered when setting up and evaluating a real-time PCR (qPCR) reaction.
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).
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.
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.
Follow example calculations to prepare and use oligonucleotide working stocks with confidence and precision.
How much do you know about TaqMan Assays? Discover some of the common misconceptions researchers have about this chemistry.
- Real-Time PCR webinars
- Genetic Analysis for All webinars
- 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?
For Research Use Only. Not for use in diagnostic procedures.