title: Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR.
abstract: Circulating (of cell-free) microRNAs (miRNAs) are released from cells into the blood stream. The amount of specific microRNAs in the circulation has been linked to a disease state and has the potential to be used as disease biomarker. A sensitive and accurate method for circulating microRNA quantification using a dye-based chemistry and droplet digital PCR technology has been recently developed. Specifically, using Locked Nucleic Acid (LNA)-based miRNA-specific primers with a green fluorescent DNA-binding dye in a compatible droplet digital PCR system it is possible to obtain the absolute quantification of specific miRNAs. Here, we describe how performing this technique to assess miRNA amount in biological fluids, such as plasma and serum, is both feasible and effective.
authors.affiliation: Department of Experimental, Diagnostic and Specialty Medicine - DIMES, University of Bologna; manuela.ferracin@unibo.it.
authors.forename: Manuela
authors.initials: M
authors.lastname: Ferracin
authors.affiliation: Department of Morphology, Surgery and Experimental Medicine, University of Ferrara.
authors.forename: Irene
authors.initials: I
authors.lastname: Salamon
authors.affiliation: Department of Morphology, Surgery and Experimental Medicine, University of Ferrara.
authors.forename: Laura
authors.initials: L
authors.lastname: Lupini
authors.affiliation: Department of Morphology, Surgery and Experimental Medicine, University of Ferrara.
authors.forename: Elena
authors.initials: E
authors.lastname: Miotto
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara.
authors.forename: Silvia
authors.initials: S
authors.lastname: Sabbioni
authors.affiliation: Department of Morphology, Surgery and Experimental Medicine and Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara; ngm@unife.it.
authors.forename: Massimo
authors.initials: M
authors.lastname: Negrini
mesh_terms: D004247:DNA; D005456:Fluorescent Dyes; D035683:MicroRNAs; D016133:Polymerase Chain Reaction
chemical_list: D005456:Fluorescent Dyes; D035683:MicroRNAs; D004247:DNA