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 language | English |
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Article number | 101515 |
Journal | STAR Protocols |
Volume | 3 |
Issue number | 3 |
Number of pages | 11 |
ISSN | 2666-1667 |
DOIs | |
Publication status | Published - 2022 |
Bibliographical note
Publisher Copyright:© 2022 The Author(s)
Keywords
- Gene Expression
- Molecular Biology