Quantification of Circulating miRNAs

The possibility of utilising circulating miRNAs to develop novel prognostic and diagnostic tests is attractive, as their expression is stable, and miRNAs can be amplified and detected with high sensitivity and specificity (Fig. 5.1). Moreover, multiple miRNAs can be assessed in a single experiment (Engelhardt 2012). However, there are several issues that need to be considered to ensure accurate quantification (Fig. 5.2). Sample preparation and data normalisation are the two most critical determinants of quantification accuracy. In this chapter, we will focus

Fig. 5.1 Experimental workflow for the quantification of circulating miRNAs

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Fig. 5.2 Considerations for the quantification of circulating miRNAs

on the challenges in the preparation of blood specimens that are used to generate plasma (supernatant of the blood collected in the presence of an anticoagulant) or serum (supernatant of coagulated blood depleted of fibrinogen).