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Enzyme enhanced quantitative determination of multiple DNA targets based on capillary electrophoresis
- Source :
-
Journal of Chromatography A . Mar2009, Vol. 1216 Issue 12, p2567-2573. 7p. - Publication Year :
- 2009
-
Abstract
- Abstract: In the current paper, enzyme enhanced simultaneous quantitative determination of multiple DNA targets based on capillary electrophoresis (CE) was described. We used three biotin-modified DNA probes, which reacted with avidin-conjugated horseradish peroxidase (avidin-HRP) conjugate to obtain the HRP labeled probes, to hybridize with three corresponding targets. The resulting mixture containing double-strand DNA (dsDNA)-HRP, excess single-strand DNA (ssDNA)-HRP and remaining avidin-HRP was separated by capillary electrophoresis, and then the system of HRP catalyzing H2O2/o-aminophenol (OAP) reaction was adopted. The catalytic product was detected with electrochemical detection. With this protocol, the limits of quantification for the hybridization assay of 21-, 39- and 80-mer DNA fragments were of 1.2×10−11, 2.4×10−11 and 3.0×10−11 M, respectively. The multiplex assay also provided good specificity without any cross-reaction. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 00219673
- Volume :
- 1216
- Issue :
- 12
- Database :
- Academic Search Index
- Journal :
- Journal of Chromatography A
- Publication Type :
- Academic Journal
- Accession number :
- 36770321
- Full Text :
- https://doi.org/10.1016/j.chroma.2009.01.019