title: Surface plasmon field-enhanced fluorescence spectroscopy in PCR product analysis by peptide nucleic acid probes.
abstract: 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.
authors.affiliation: Max-Planck-Institute for Polymer Research, Ackermannweg 10, 55128, Mainz, Germany.
authors.forename: Danfeng
authors.initials: D
authors.lastname: Yao
authors.forename: Fang
authors.initials: F
authors.lastname: Yu
authors.forename: Junyoung
authors.initials: J
authors.lastname: Kim
authors.forename: Judith
authors.initials: J
authors.lastname: Scholz
authors.forename: Peter E
authors.initials: PE
authors.lastname: Nielsen
authors.forename: Eva-Kathrin
authors.initials: EK
authors.lastname: Sinner
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Knoll
mesh_terms: D016375:Antisense Elements (Genetics); D009693:Nucleic Acid Hybridization / Q000379:methods*; D015341:Nucleic Acid Probes / Q000737:chemistry*; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D013050:Spectrometry, Fluorescence / Q000379:methods*; D020349:Surface Plasmon Resonance
chemical_list: D016375:Antisense Elements (Genetics); D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids