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.