Quantification of global microRNA abundance by selective isotachophoresis

Alexandre Persat, Raghu R. Chivukula, Joshua T. Mendell, Juan G. Santiago

Research output: Contribution to journalArticle

Abstract

We here present and demonstrate a novel technique based on isotachophoresis (ITP) for the quantification of global microRNA (miRNA) abundance in total RNA. We leverage the selectivity of ITP to concentrate miRNA and exclude longer RNA molecules from the focused zone. We designed a novel ITP strategy where we initially establish three contiguous zones of sieving polymer, electrolyte, and denaturant concentrations. This allows for successive preconcentration, selection, and detection of miRNA. We optimized chemistry in each zone for high sensitivity and exquisite selectivity for miRNA. This technique allows for the measurement of the total miRNA content in a sample and its comparison between different cell types and tissues. We demonstrated and validated the efficacy of this technique by comparing global miRNA abundance in subconfluent and confluent cell cultures.

Original languageEnglish (US)
Pages (from-to)9631-9635
Number of pages5
JournalAnalytical Chemistry
Volume82
Issue number23
DOIs
StatePublished - Dec 1 2010

Fingerprint

MicroRNAs
RNA
Cell culture
Electrolytes
Isotachophoresis
Polymers
Tissue
Molecules

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Persat, A., Chivukula, R. R., Mendell, J. T., & Santiago, J. G. (2010). Quantification of global microRNA abundance by selective isotachophoresis. Analytical Chemistry, 82(23), 9631-9635. https://doi.org/10.1021/ac102496m

Quantification of global microRNA abundance by selective isotachophoresis. / Persat, Alexandre; Chivukula, Raghu R.; Mendell, Joshua T.; Santiago, Juan G.

In: Analytical Chemistry, Vol. 82, No. 23, 01.12.2010, p. 9631-9635.

Research output: Contribution to journalArticle

Persat, A, Chivukula, RR, Mendell, JT & Santiago, JG 2010, 'Quantification of global microRNA abundance by selective isotachophoresis', Analytical Chemistry, vol. 82, no. 23, pp. 9631-9635. https://doi.org/10.1021/ac102496m
Persat, Alexandre ; Chivukula, Raghu R. ; Mendell, Joshua T. ; Santiago, Juan G. / Quantification of global microRNA abundance by selective isotachophoresis. In: Analytical Chemistry. 2010 ; Vol. 82, No. 23. pp. 9631-9635.
@article{4c344661b0cd4c3b9553e21205752db7,
title = "Quantification of global microRNA abundance by selective isotachophoresis",
abstract = "We here present and demonstrate a novel technique based on isotachophoresis (ITP) for the quantification of global microRNA (miRNA) abundance in total RNA. We leverage the selectivity of ITP to concentrate miRNA and exclude longer RNA molecules from the focused zone. We designed a novel ITP strategy where we initially establish three contiguous zones of sieving polymer, electrolyte, and denaturant concentrations. This allows for successive preconcentration, selection, and detection of miRNA. We optimized chemistry in each zone for high sensitivity and exquisite selectivity for miRNA. This technique allows for the measurement of the total miRNA content in a sample and its comparison between different cell types and tissues. We demonstrated and validated the efficacy of this technique by comparing global miRNA abundance in subconfluent and confluent cell cultures.",
author = "Alexandre Persat and Chivukula, {Raghu R.} and Mendell, {Joshua T.} and Santiago, {Juan G.}",
year = "2010",
month = "12",
day = "1",
doi = "10.1021/ac102496m",
language = "English (US)",
volume = "82",
pages = "9631--9635",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "23",

}

TY - JOUR

T1 - Quantification of global microRNA abundance by selective isotachophoresis

AU - Persat, Alexandre

AU - Chivukula, Raghu R.

AU - Mendell, Joshua T.

AU - Santiago, Juan G.

PY - 2010/12/1

Y1 - 2010/12/1

N2 - We here present and demonstrate a novel technique based on isotachophoresis (ITP) for the quantification of global microRNA (miRNA) abundance in total RNA. We leverage the selectivity of ITP to concentrate miRNA and exclude longer RNA molecules from the focused zone. We designed a novel ITP strategy where we initially establish three contiguous zones of sieving polymer, electrolyte, and denaturant concentrations. This allows for successive preconcentration, selection, and detection of miRNA. We optimized chemistry in each zone for high sensitivity and exquisite selectivity for miRNA. This technique allows for the measurement of the total miRNA content in a sample and its comparison between different cell types and tissues. We demonstrated and validated the efficacy of this technique by comparing global miRNA abundance in subconfluent and confluent cell cultures.

AB - We here present and demonstrate a novel technique based on isotachophoresis (ITP) for the quantification of global microRNA (miRNA) abundance in total RNA. We leverage the selectivity of ITP to concentrate miRNA and exclude longer RNA molecules from the focused zone. We designed a novel ITP strategy where we initially establish three contiguous zones of sieving polymer, electrolyte, and denaturant concentrations. This allows for successive preconcentration, selection, and detection of miRNA. We optimized chemistry in each zone for high sensitivity and exquisite selectivity for miRNA. This technique allows for the measurement of the total miRNA content in a sample and its comparison between different cell types and tissues. We demonstrated and validated the efficacy of this technique by comparing global miRNA abundance in subconfluent and confluent cell cultures.

UR - http://www.scopus.com/inward/record.url?scp=78649706595&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649706595&partnerID=8YFLogxK

U2 - 10.1021/ac102496m

DO - 10.1021/ac102496m

M3 - Article

C2 - 21062022

AN - SCOPUS:78649706595

VL - 82

SP - 9631

EP - 9635

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 23

ER -