Multiplexed microRNA detection by capillary electrophoresis with laser-induced fluorescence

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Please use this identifier to cite or link to this item: http://140.128.103.80:8080/handle/310901/23116

Title:	Multiplexed microRNA detection by capillary electrophoresis with laser-induced fluorescence
Authors:	Jiang, R.-M., Chang, Y.-S., Chen, S.-J., Chen, J.-H., Chen, H.-C., Chang, P.-L.
Contributors:	Department of Chemistry, Tunghai University
Keywords:	Capillary electrophoresis;Laser-induced fluorescence;Poly(ethylene) oxide;RNA;Separation
Date:	2011
Issue Date:	2013-06-04T08:22:55Z (UTC)
Abstract:	In this study, we developed a novel assay that simultaneously detects multiple miRNAs (microRNAs) within a single capillary by combining a tandem adenosine-tailed DNA bridge-assisted splinted ligation with denaturing capillary gel electrophoresis with laser-induced fluorescence. This proposed method not only represents a significant improvement in resolution but also allows for the detection of multiple miRNAs within a single capillary based on the length differences of specified target bridge DNA. The assay's linear range covers three orders of magnitude (1.0. nM to 1.0. pM) with a limit of detection (S/N. =3) as low as 190. fM (2.5. zmol). Five miRNAs of Epstein-Barr virus (EBV) were also detected in EBV-infected nasopharyngeal carcinoma cells, while they did not appear in non-virus infected cells. Moreover, the electropherogram indicated that the screening of isomiRs (isomer of miRNA) of BART2 by CE-LIF is feasible by our proposed method. The developed electrophoresis-based method for miRNA detection is fast, amplification-free, multiplexed and cost-effective, making it potentially applicable to large-scale screening of isomiRs. ? 2011 Elsevier B.V.
Relation:	Journal of Chromatography A 1218 (18) , pp. 2604-2610
Appears in Collections:	[Department of Chemistry ] Periodical Articles

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