Protocol for qPCR analysis that corrects for cDNA amplification efficiency

Mads V. Damgaard*, Jonas T. Treebak

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

17 Citations (Scopus)
26 Downloads (Pure)

Abstract

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
Volume3
Issue number3
Number of pages11
ISSN2666-1667
DOIs
Publication statusPublished - 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s)

Keywords

  • Gene Expression
  • Molecular Biology

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