TIPS & TECHNIQUES qPCR Basics A Amplified product is visualized by agarose gel after it is produced (thus the end-point designation) Does not precisely measure the starting DNA or RNA Less expensive; no special instruments required Basic molecular biology technique to Help with Successful Real-time Amplification n increasingly popular amplification technique is real-time or quantitative PCR (qPCR). Here is a brief comparison of endpoint PCR and qPCR, plus an exploration of how qPCR works, and things to consider when setting up and performing qPCR experiments to get the best results. Comparison of Traditional Endpoint PCR and qPCR Endpoint PCR qPCR Amplified product is visualized as it is produced (in real time) using a detection instrument Allows you to measure how many copies of DNA or RNA you started with (q = quantitative) More expensive; requires special instrumentation Requires slightly more technical ability qPCR can be used to answer the same experimental questions as endpoint PCR, including detecting polymorphisms, amplifying low-abundance sequences for cloning or analysis, and pathogen detection. In addition, with qPCR you can see amplification in real time and detect the number of copies in the starting material, which makes it possible to quantitate gene expression, measure the amount of DNA damage or quantitate viral load in a sample. enews | November 2015 Pagina 9
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Labnet Oy - Promega's Monthly online Magazine - November 2015 edition Lees publicatie 101Home