Expression Stability Analysis of Candidate References for Normalization of RT-qPCR Data Using RefSeeker R package
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When performing expression analysis either for coding RNA (e.g., mRNA) or non-coding RNA (e.g., miRNA), reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a widely used method. To normalize these data, one or more stable endogenous references must be identified. RefFinder is an online web-based tool using four almost universally used algorithms for assessing candidate endogenous references—delta-Ct, BestKeeper, geNorm, and Normfinder. However, the online interface is presently cumbersome and time consuming. We developed an R package, RefSeeker, which performs easy and straightforward RefFinder analysis by enabling raw data import and calculation of stability from each of the algorithms and provides data output tools to create graphs and tables. This protocol uses RefSeeker R package for fast and simple RefFinder stability analysis.
Original language | English |
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Article number | e4801 |
Journal | Bio-protocol |
Volume | 13 |
Issue number | 17 |
Number of pages | 14 |
ISSN | 2331-8325 |
DOIs | |
Publication status | Published - 2023 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:
© 2023 The Authors; exclusive licensee Bio-protocol LLC.
- Coding and non-coding RNA expression, Expression stability, Normalization, R package, RefFinder, RT-qPCR
Research areas
ID: 396719549