Abstract
microRNAs (miRNAs), short noncoding RNAs, are posttranscriptional negative regulators, with extracellular circulating miRNAs considered promising biomarkers of various diseases. The relative quantification of miRNA transcripts requires an endogenous reference gene for data normalization, which is the critical step in this process. In the present chapter, the means of normalization and methods for the selection of suitable reference gene(s) are discussed, with a key finding being how common reference genes such as RNU6 and miR-16 can cause bias in data interpretation if used without proper validation.
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Abbreviations
- 18S:
-
Ribosomal RNA 18S
- 5S:
-
Ribosomal RNA 5S
- ACT:
-
Actin
- Cq:
-
Quantitation cycle
- FFPE:
-
Formalin-fixed paraffin-embedded
- GAPDH:
-
Glyceraldehyde 3-phosphate dehydrogenase
- LNA:
-
Locked nucleic acid
- MIQE:
-
Minimum information for publication of quantitative real-time PCR experiments
- miRNA:
-
microRNA
- qPCR:
-
Quantitative real-time polymerase chain reaction
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Matoušková, P. (2019). MicroRNAs and Reference Gene Methodology. In: Patel, V., Preedy, V. (eds) Handbook of Nutrition, Diet, and Epigenetics. Springer, Cham. https://doi.org/10.1007/978-3-319-55530-0_34
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