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.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 101515 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 3 |
| Udgave nummer | 3 |
| Antal sider | 11 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
This work was supported by a PhD scholarship to M.V.D. from the Danish Diabetes Academy , which is funded by the Novo Nordisk Foundation ( NNF17SA0031406 ). Further financial support was provided by Novo Nordisk Foundation Center for Basic Metabolic Research (CBMR). CBMR is an independent Research Center at the University of Copenhagen , which is partially funded by an unrestricted donation from the Novo Nordisk Foundation ( NNF18CC0034900 ). The graphical abstract was created using assets from BioRender.com.
Publisher Copyright:
© 2022 The Author(s)
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