Optimization techniques for miRNA expression in low frequency immune cell populations

Victoria G. Lyons, Natalie L. Payne, Claire E. McCoy

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

In this chapter we outline a RNA extraction method for very low immune cell populations isolated from the central nervous system of mice undergoing experimental autoimmune encephalomyelitis. We compare various normalization and quantification techniques to examine miRNA expression. Our data highlight that employing a mean normalization procedure using a number of well-selected housekeeping miRNA genes, followed by absolute quantification with a standard curve generated from a commercial miRNA oligo, gave the most robust and reproducible miRNA expression results.

Original languageEnglish
Title of host publicationInflammation and Cancer
Subtitle of host publicationMethods and Protocols
EditorsBrendan J. Jenkins
Place of PublicationNew York, NY
PublisherHumana Press
Pages237-256
Number of pages20
ISBN (Electronic)9781493975686
ISBN (Print)9781493975679
DOIs
Publication statusPublished - 1 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1725
ISSN (Print)1064-3745

Keywords

  • Absolute quantification
  • Experimental autoimmune encephalomyelitis
  • Immune cells
  • Mean normalization
  • microRNA
  • Relative quantification
  • RT-PCR
  • Standard curve

Cite this

Lyons, V. G., Payne, N. L., & McCoy, C. E. (2018). Optimization techniques for miRNA expression in low frequency immune cell populations. In B. J. Jenkins (Ed.), Inflammation and Cancer: Methods and Protocols (pp. 237-256). (Methods in Molecular Biology; Vol. 1725). Humana Press. https://doi.org/10.1007/978-1-4939-7568-6_19