Reliable qPCR results depend not only on features of your region of interest (you must consider transcript variants, exon boundaries, and SNPs), but also on several experimental parameters. Therefore, it’s important to validate your assays, even those that have been used in previous publications. If you’re not validating under your own conditions, you could end up wasting valuable time and precious samples.
This special edition Real-time qPCR guide: Part 2—assay validation and data analysis covers critical variables for accurate and efficient assays. Understanding these details will allow you to design experiments that generate reliable and interpretable data.
(if you missed receiving our guide for assay design, access it here: Real-time qPCR guide: Part 1—assay design)