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

被引：8

作者：

Yamamura, Shohei ^{[1
]}

Yatsushiro, Shouki ^{[1
]}

Yamaguchi, Yuka ^{[1
]}

Abe, Kaori ^{[1
]}

Shinohara, Yasuo ^{[2
]}

Kataoka, Masatoshi ^{[1
]}

机构：

[1] Natl Inst Adv Ind Sci & Technol, Hlth Res Inst, Takamatsu, Kagawa 7610395, Japan

[2] Univ Tokushima, Inst Genome Res, Div Prot Express, Tokushima 7708503, Japan

来源：

SENSORS | 2012年 / 12卷 / 06期

关键词：

microRNA; RNase protection assay; microchip electrophoresis; MICRORNA; CANCER;

D O I：

10.3390/s120607576

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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.

引用

页码：7576 / 7586

页数：11

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