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). New York, NY: Humana Press. https://doi.org/10.1007/978-1-4939-7568-6_19
Lyons, Victoria G. ; Payne, Natalie L. ; McCoy, Claire E. / Optimization techniques for miRNA expression in low frequency immune cell populations. Inflammation and Cancer: Methods and Protocols. editor / Brendan J. Jenkins. New York, NY : Humana Press, 2018. pp. 237-256 (Methods in Molecular Biology).
@inbook{03853569e9554a809fadd0b7515b43fe,
title = "Optimization techniques for miRNA expression in low frequency immune cell populations",
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.",
keywords = "Absolute quantification, Experimental autoimmune encephalomyelitis, Immune cells, Mean normalization, microRNA, Relative quantification, RT-PCR, Standard curve",
author = "Lyons, {Victoria G.} and Payne, {Natalie L.} and McCoy, {Claire E.}",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-7568-6_19",
language = "English",
isbn = "9781493975679",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "237--256",
editor = "Jenkins, {Brendan J.}",
booktitle = "Inflammation and Cancer",
address = "United States of America",

}

Lyons, VG, Payne, NL & McCoy, CE 2018, Optimization techniques for miRNA expression in low frequency immune cell populations. in BJ Jenkins (ed.), Inflammation and Cancer: Methods and Protocols. Methods in Molecular Biology, vol. 1725, Humana Press, New York, NY, pp. 237-256. https://doi.org/10.1007/978-1-4939-7568-6_19

Optimization techniques for miRNA expression in low frequency immune cell populations. / Lyons, Victoria G.; Payne, Natalie L.; McCoy, Claire E.

Inflammation and Cancer: Methods and Protocols. ed. / Brendan J. Jenkins. New York, NY : Humana Press, 2018. p. 237-256 (Methods in Molecular Biology; Vol. 1725).

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

TY - CHAP

T1 - Optimization techniques for miRNA expression in low frequency immune cell populations

AU - Lyons, Victoria G.

AU - Payne, Natalie L.

AU - McCoy, Claire E.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

KW - Absolute quantification

KW - Experimental autoimmune encephalomyelitis

KW - Immune cells

KW - Mean normalization

KW - microRNA

KW - Relative quantification

KW - RT-PCR

KW - Standard curve

UR - http://www.scopus.com/inward/record.url?scp=85040664511&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-7568-6_19

DO - 10.1007/978-1-4939-7568-6_19

M3 - Chapter (Book)

SN - 9781493975679

T3 - Methods in Molecular Biology

SP - 237

EP - 256

BT - Inflammation and Cancer

A2 - Jenkins, Brendan J.

PB - Humana Press

CY - New York, NY

ER -

Lyons VG, Payne NL, McCoy CE. Optimization techniques for miRNA expression in low frequency immune cell populations. In Jenkins BJ, editor, Inflammation and Cancer: Methods and Protocols. New York, NY: Humana Press. 2018. p. 237-256. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7568-6_19