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P36 VOLTAMETRIC DETECTION OF DNA DELETION ON PENCIL ELECTRODE
Haroniková, Lucia ; Špaček, Jan ; Fojta, Miroslav
In this work, we present a new qualitative approach of detection of PCR products using electrochemistry on pencil electrodes. PCR products with an incorporated biotin-labeled dNTP (dCTP in this study) are detected via conjugated streptavidinealkaline phospatase. 1-Naphthyl phosphate (1-NP), which is dephosphorylated by alkaline phosphatase to release 1-naphthol, was used as a substrate in electrochemical detection of the PCR product. Voltammetric measurement on disposable pencil electrode is a very cheap and easy to use method. The system was optimized for plasmid DNA PCR product with potential to detect human genome DNA products and possible application in gene deletion monitoring.
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Examples of Using of Electrochemical Detection at Pencil Graphite Electrode with Enzymatic Labeling for Analysis of Nucleotide Sequence
Plucnara, Medard ; Haroniková, Lucia ; Špaček, Jan ; Havran, Luděk ; Horáková, Petra ; Pivoňková, Hana ; Ecsin, E. ; Erdem, A. ; Fojta, Miroslav
Many examples of utilization of enzymatic labeling for DNA sequence analysis has been described in literature so far. Some of them involve hybridization with complementary biotinylated probe, while others use incorporation of biotinylated nucleotides into DNA strand by DNA polymerases. Common approach is then binding of streptavidine-enzyme conjugates to biotin tags and incubation with substrate, which is converted to detectable product. Here, two recent applications using this principle are described for the detection of PCR amplicons and for SNP typing. Both techniques are combined with detection at pencil graphite electrodes.
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