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Guidelines for a Successful qPCR Master Mix Comparison

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Abstract

Real-time quantitative PCR (qPCR) is a powerful tool to detect and quantify nucleic acids. By incorporating fluorescently labeled probes or fluorescent double-stranded DNA (dsDNA)-binding dyes into the PCR, product formation can be monitored following each PCR cycle. GoTaq® qPCR Master Mixes for dye-based or probe-based detection are optimized for fast and reproducible qPCR assays. In this guide, we outline some of the most important considerations for comparing the performance of your assay using either GoTaq® qPCR Master Mix with BRYT® Green Dye or GoTaq® Probe qPCR Master Mix to that of your current qPCR reagent. Testing previously-optimized qPCR assays with a new qPCR master mix requires careful experimental design to compare several factors of reagent performance including assay specificity, repeatability, linearity, sensitivity and efficiency. Each of these qPCR assay performance indicators will be discussed with respect to how the qPCR master mix comparison experiments should be performed as well as how to analyze the data.

Sarah Teter, PhD and Leta Steffen, PhD

Promega Corporation

Publication Date: 8/16

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How to Cite This Article

Teter, S. and Steffen, L. Guidelines for a Successful qPCR Master Mix Comparison. [Internet] 8/16. [cited: year, month, date]. Available from: http://www.promega.com/resources/pubhub/guidelines-for-a-comparison-of-qpcr-reagent-performance/

Teter, S. and Steffen, L. Guidelines for a Successful qPCR Master Mix Comparison. Promega Corporation Web site. http://www.promega.com/resources/pubhub/guidelines-for-a-comparison-of-qpcr-reagent-performance/ Updated 8/16. Accessed Month Day, Year.

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