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Quantitative PCR (a.k.a. qPCR or real-time PCR) is one of the most commonly used techniques in molecular life sciences. It is used for multiple applications including gene expression, SNP genotyping, and copy number variation. However, knowing how to achieve the best results is not always straightforward. In this webinar on 5′ nuclease (primer- and probe-based) qPCR assay design and experimental setup, senior scientific applications specialist Dr Erik Wendlandt provides guidelines for using design parameters such as Tm, GC content, amplicon size, and primer and probe location. Dr Wendlandt describes several free online tools, including BLAST, that can aid in primer/probe design and analysis. General considerations for droplet digital PCR (ddPCR) are included.