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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....
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Molecular Diversity Preservation International (MDPI)
2012
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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. |
format | Online Article Text |
id | pubmed-3435990 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Molecular Diversity Preservation International (MDPI) |
record_format | MEDLINE/PubMed |
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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