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Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe

The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA....

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Detalles Bibliográficos
Autores principales: Yamamura, Shohei, Yatsushiro, Shouki, Yamaguchi, Yuka, Abe, Kaori, Shinohara, Yasuo, Kataoka, Masatoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435990/
https://www.ncbi.nlm.nih.gov/pubmed/22969361
http://dx.doi.org/10.3390/s120607576
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author Yamamura, Shohei
Yatsushiro, Shouki
Yamaguchi, Yuka
Abe, Kaori
Shinohara, Yasuo
Kataoka, Masatoshi
author_facet Yamamura, Shohei
Yatsushiro, Shouki
Yamaguchi, Yuka
Abe, Kaori
Shinohara, Yasuo
Kataoka, Masatoshi
author_sort Yamamura, Shohei
collection PubMed
description The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA. The fluorescence intensity corresponding to the protected RNA fragment increased in a dose-dependent manner with respect to the complementary-strand RNA. More highly sensitive detection of miRNA by microchip electrophoresis than by conventional method using fluorescence-labeled riboprobe could be obtained in 180 s. An obvious increase in miR-222 expression induced by nerve growth factor in PC12 cells could be observed. These results clearly indicate the potential of microchip electrophoresis for the analysis of miRNA using RNase protection assay with a fluorescence-labeled riboprobe.
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spelling pubmed-34359902012-09-11 Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe Yamamura, Shohei Yatsushiro, Shouki Yamaguchi, Yuka Abe, Kaori Shinohara, Yasuo Kataoka, Masatoshi Sensors (Basel) Article The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA. The fluorescence intensity corresponding to the protected RNA fragment increased in a dose-dependent manner with respect to the complementary-strand RNA. More highly sensitive detection of miRNA by microchip electrophoresis than by conventional method using fluorescence-labeled riboprobe could be obtained in 180 s. An obvious increase in miR-222 expression induced by nerve growth factor in PC12 cells could be observed. These results clearly indicate the potential of microchip electrophoresis for the analysis of miRNA using RNase protection assay with a fluorescence-labeled riboprobe. Molecular Diversity Preservation International (MDPI) 2012-06-07 /pmc/articles/PMC3435990/ /pubmed/22969361 http://dx.doi.org/10.3390/s120607576 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Yamamura, Shohei
Yatsushiro, Shouki
Yamaguchi, Yuka
Abe, Kaori
Shinohara, Yasuo
Kataoka, Masatoshi
Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title_full Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title_fullStr Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title_full_unstemmed Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title_short Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe
title_sort detection of mirna in cell cultures by using microchip electrophoresis with a fluorescence-labeled riboprobe
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435990/
https://www.ncbi.nlm.nih.gov/pubmed/22969361
http://dx.doi.org/10.3390/s120607576
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