Normalization panels for RT-qPCR experiments on circulating miRNAs

Over the past decade, miRNAs emerged as useful biomarkers. Quantitative polymerase chain reaction
(qPCR) is the most commonly used method to detect specifi c miRNAs, but to obtain reliable results,
proper normalization is a necessity. The three most commonly used strategies for normalizing miRNA
levels are:

  1. normalization to the geometrical mean of all detected miRNAs,
  2. normalization to a single endogenous control (for example RNU6B or miR-16), and
  3. the use of a spike-in.

Pros and cons have been described for all these normalization methods, but none of them seems
ideal for universal use in qPCR experiments. Specific miRNAs that can be used for normalization have
also been reported, but as their use is limited to the diseased population in which they were studied,
they also do not provide a universal normalization method.


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