Detection of microRNAs Using Chip-Based QuantStudio 3D Digital PCR

Methods Mol Biol. 2017:1580:239-247. doi: 10.1007/978-1-4939-6866-4_16.

Abstract

Digital PCR (dPCR) is an innovative approach for detection and quantification of nucleic acid that offers an alternative method to conventional real-time quantitative PCR for absolute quantification. dPCR is a highly precise and sensitive technique that does not require a standard reference, making it a suitable method for the detection of microRNAs. The potential of these small noncoding RNA as biomarkers is on the rise, especially due to their presence in body fluids, making them easily accessible. Nevertheless, the problem of lack of consensus regarding an optimal method for miRNAs normalization compromises their use. Here, we describe an innovative method for the absolute quantification of miRNAs across different types of biological samples using a chip-based platform, the QuantStudio 3D digital PCR.

Keywords: Absolute quantification; Digital PCR; miRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Equipment Design
  • Humans
  • MicroRNAs / genetics*
  • MicroRNAs / isolation & purification
  • Oligonucleotide Array Sequence Analysis / instrumentation*
  • Oligonucleotide Array Sequence Analysis / methods
  • Polymerase Chain Reaction / instrumentation*
  • Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction / instrumentation
  • Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

  • MicroRNAs