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Melt-curve analysis for improved intercalating dye qPCR

The benefits and limitations of melt-curve analysis are presented in this IDT-hosted webinar. Learn how PrimeTime Predesigned qPCR Primer Assays are compatible with intercalating dyes, and how they can be transitioned to probe-based assays.

Performing qPCR using intercalating dyes, such as SYBR® dyes, involves use of just 2 amplification primers to provide assay specificity. Because fluorescence in intercalating dye assays is based on the production of any double-stranded DNA, off-target amplification can generate false-positive signals, compromising assay data. To address this problem, researchers use melt-curve analysis to assess whether their assays have produced single, specific products.

In April 2015, IDT hosted a webinar, Understanding Melt Curves for Improved SYBR Green Assay Analysis and Troubleshooting, that covered several aspects of melt curve analysis. Now available as a recording, this webinar features our resident expert, Nick Downey, PhD, demonstrating some of the benefits and limitations of melt-curve analysis through clear examples. The presentation, which you can watch here, also provides an overview of the uMeltSM software for guiding assay design.

Published Apr 19, 2015