PMID: 11332896May 3, 2001Paper

Fluorescence-labelled DNA probes to detect complementary sequences in homogeneous media

Journal of Photochemistry and Photobiology. B, Biology
E TalaveraJ M Alvarez-Pez

Abstract

Sensitive, safe and easy-to-use probes for the detection of nucleic acids are urgently called for. To this end we are in the process of developing a fluorescence-based technique to work in homogeneous assay media. We have examined pyrene and fluorescein as fluorescent labels for natural DNA probes. A fraction of the cytosine residues of a single-stranded cDNA was randomly labelled with either pyrene or fluorescein using the bisulfite-catalyzed diamine reaction. Both fluorophores showed fluorescence quenching when the labelled probe was hybridized with its complementary strand and we describe the changes in steady-state fluorescence intensity that occurred upon hybridization. Our results demonstrate that pyrene quenching is more efficient than fluorescein quenching and thus pyrene-labelled probes are more sensitive for detecting and quantifying DNA from natural sources.

References

Feb 15, 1990·Analytical Biochemistry·M Stürzl, W K Roth
Aug 24, 1970·Biochemical and Biophysical Research Communications·R Shapiro, J M Weisgras
Mar 31, 1981·Biochemistry·B M LiuJ Mestecky
Jul 28, 1995·Science·H NeumannH Wekerle

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Citations

Dec 21, 2007·The Journal of Physical Chemistry. a·Luis CrovettoJose M Alvarez-Pez
Jul 22, 2008·The Journal of Physical Chemistry. B·Luis CrovettoEva M Talavera
Sep 22, 2005·Journal of the American Chemical Society·Patrick J HrdlickaJesper Wengel

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