Published online 14 December 2004
Nucleic Acids Research, Vol. 32 No. 22 © Oxford University Press 2004; all rights reserved
Surface plasmon field-enhanced fluorescence spectroscopy in PCR product analysis by peptide nucleic acid probes
1 Max-Planck-Institute for Polymer Research, Ackermannweg 10, 55128, Mainz, Germany, 2 Max-Planck-Institute of Biochemistry, Am Klopferspitz 18A, 82152, Munich, Germany and 3 Department of Biochemistry B, The Panum Institute, IMBG, Blegdamsvej 3c, DK-2200 Copenhagen N, Denmark
* To whom correspondence should be addressed. Tel: +49 6131 379 160; Fax: +49 6131 379 360; Email: knoll{at}mpip-mainz.mpg.de
Received June 10, 2004; Revised September 15, 2004; Accepted November 24, 2004
Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) was recently developed for PCR product analysis, which allowed for real-time monitoring of hybridization processes and for the detection of trace amounts of PCR products, with a detection limit of 100 fmol on the peptide nucleic acid (PNA) probe surface, and 500 fmol on the DNA probe surface. By selectively labeling the strands of PCR-amplified DNA, it was shown that the heat denaturation process in combination with the application of low-salt condition substantially reduced the interference from the antisense strands and thus simplified the surface hybridization. Furthermore, SPFS was demonstrated to be capable of quantitatively discriminating the difference induced by single nucleotide substitution, even within one minute of contact time.