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
收藏
页码:7576 / 7586
页数:11
相关论文
共 50 条
  • [1] Electrophoresis on a microfluidic chip for analysis of fluorescence-labeled human rhinovirus
    Kolivoska, Viliarn
    Weiss, Victor U.
    Kremser, Leopold
    Gas, Bohuslav
    Blaas, Dieter
    Kenndler, Ernst
    ELECTROPHORESIS, 2007, 28 (24) : 4734 - 4740
  • [2] Sensitive and specific detection of miRNA using an isothermal exponential amplification method using fluorescence-labeled LNA/DNA chimera primers
    Huang, Jun-Fu
    Zhao, Na
    Xu, Han-Qing
    Xia, Han
    Wei, Kun
    Fu, Wei-Ling
    Huang, Qing
    ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2016, 408 (26) : 7437 - 7446
  • [3] Sensitive and specific detection of miRNA using an isothermal exponential amplification method using fluorescence-labeled LNA/DNA chimera primers
    Jun-Fu Huang
    Na Zhao
    Han-Qing Xu
    Han Xia
    Kun Wei
    Wei-Ling Fu
    Qing Huang
    Analytical and Bioanalytical Chemistry, 2016, 408 : 7437 - 7446
  • [4] Validation of chimerism testing using fluorescence-labeled multiplex primers with analysis by capillary electrophoresis
    Smith, AG
    McFarland, C
    Carlson, T
    Schmitt, F
    Hansen, J
    HUMAN IMMUNOLOGY, 2004, 65 : S116 - S116
  • [5] Detection of single oxygen molecules with fluorescence-labeled hemocyanins
    Erker, W
    Sdorra, S
    Basché, T
    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (42) : 14532 - 14533
  • [6] Detection of single oxygen molecules with fluorescence-labeled hemocyanins
    1600, American Chemical Society, Columbus, United States (127):
  • [7] IMMUNOASSAY USING HPLAC AND FLUORESCENCE-LABELED ANTIBODIES
    YOSHIKAWA, T
    TERASHIMA, M
    KATOH, S
    JOURNAL OF FERMENTATION AND BIOENGINEERING, 1995, 80 (02): : 200 - 203
  • [8] Dual fluorescence and electrochemical detection on an electrophoresis microchip
    Lapos, JA
    Manica, DP
    Ewing, AG
    ANALYTICAL CHEMISTRY, 2002, 74 (14) : 3348 - 3353
  • [9] Detection of hyphal fusion in filamentous fungi using differently fluorescence-labeled histones
    Rech, Christine
    Engh, Ines
    Kueck, Ulrich
    CURRENT GENETICS, 2007, 52 (5-6) : 259 - 266
  • [10] Detection of hyphal fusion in filamentous fungi using differently fluorescence-labeled histones
    Christine Rech
    Ines Engh
    Ulrich Kück
    Current Genetics, 2007, 52 : 259 - 266