Publication detail

Utilization of Electrochemical Activity of 7-Deazaguanine in the Monitoring of PCR Amplification of G-rich DNA Strands

Authors: Fojtová Miloslava | Horáková Petra | Pivoňková Hana | Vychodilová Zdenka | Fojta Miroslav

Year: 2010

Type of publication: článek ve sborníku

Name of source: Sborník přednášek mezinárodní odborné konference XXX. Moderní elektrochemické metody

Publisher name: BEST Servis

Place: Ústí nad Labem

Page from-to: 50-54

Titles:

Language	Name	Abstract	Keywords
cze	Využití elektrochemické aktivity 7-deazaguaninu při monitorování PCR amplifikace G-bohatých řetězců DNA	7-deazaguanin je analog guaninu, který lze sekvenčně specificky inkorporovat do DNA. Inkorporace 7-deazaguaninu do PCR amplikonů byla sledována pomocí voltametrické analýzy una hlíkových elektrodách.	7-Deazaguanin; elektrochemická analýza; značení DNA; PCR; PCR v reálném čase
eng	Utilization of Electrochemical Activity of 7-Deazaguanine in the Monitoring of PCR Amplification of G-rich DNA Strands	7-Deazaguanine (G) is an analogue of guanine incorporable sequence-specifically into DNA but incapable of forming Hoogsteen base pairs. Reduced tendency of G to form alternative DNA structures is utilized in PCR amplification of guanine-rich nucleotide sequences. G* is electrochemically active, being oxidized at significantly lower potentials, compared to guanine and all other nucleobases. We show that incorporation of G* into PCR amplicons can easily be monitored by voltammetric analysis of the modified DNA at carbon electrodes. Densely G-modified DNA fragments exhibit quenching of fluorescence of SYBR Green I dye, commonly used for gel staining and real time PCR applications, the electrochemical detection provides G-specific signal suitable for the quantification of the amplified DNA as well as for the determination of the DNA modification extent.	7-Deazaguanine; Electrochemical analysis; DNA labeling; PCR; Real-time PCR

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