Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

被引：8

作者：

Ferracin, Manuela ^{[1
]}

Salamon, Irene ^{[2
]}

Lupini, Laura ^{[2
]}

Miotto, Elena ^{[2
]}

Sabbioni, Silvia ^{[3
]}

Negrini, Massimo ^{[2
,4
]}

机构：

[1] Univ Bologna, Dept Expt Diagnost & Specialty Med DIMES, I-40126 Bologna, Italy

[2] Univ Ferrara, Dept Morphol Surg & Expt Med, I-44100 Ferrara, Italy

[3] Univ Ferrara, Dept Life Sci & Biotechnol, I-44100 Ferrara, Italy

[4] Univ Ferrara, Lab Technol Adv Therapies LTTA, I-44100 Ferrara, Italy

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2016年 / 112期

关键词：

Bioengineering; Issue; 112; MicroRNA; serum; plasma; droplet digital PCR; diagnostics; cancer; biomarker; BIOMARKERS; MIRNAS;

D O I：

10.3791/54102

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：6

共 16 条

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