Protocol for qPCR analysis that corrects for cDNA amplification efficiency

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This protocol presents a variation on the 2-ΔΔCt technique for qPCR analysis. Our approach requires the inclusion of a standard curve on each qPCR plate, and like the 2-ΔΔCt technique, is dependent on the stability of housekeeping gene expression. However, unlike the 2-ΔΔCt technique, our approach corrects for imperfect cDNA amplification efficiency and allows for the use of multiple housekeeping genes. Collectively, this approach enhances analytical accuracy and thereby reduces the type I and II statistical errors in the generated data.

Original languageEnglish
Article number101515
JournalSTAR Protocols
Issue number3
Number of pages11
Publication statusPublished - 2022

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© 2022 The Author(s)

    Research areas

  • Gene Expression, Molecular Biology

ID: 314837572