title: New tools for oligonucleotide fingerprinting.
abstract: Oligonucleotide fingerprinting is an attractive, high-throughput complement to tag sequencing methods to determine the spectrum and abundance of genes in cDNA libraries. This method currently relies on the sequential hybridizations of short, radioactively labeled DNA oligonucleotides to clone arrays. Here, we describe a new environment that substantially improves this technology. Fluorescently labeled peptide nucleic acid (PNA) oligonucleotides are used as hybridization probes. Hybridization results are recorded with a large-field, high-resolution laser scanner developed for this purpose. Automated image analysis allows easy handling of large numbers of hybridization images. Signal interference effects, which limit the gridding density in the radioactive approach, are strongly reduced. The sensitivity of the fluorescence detection demonstrated permits the convenient use of nylon membranes. Hybridization data quality is improved, and its generation is substantially accelerated, simplified, and less expensive.
authors.affiliation: Max-Planck-Institute for Molecular Genetics, Berlin, Germany. guer@molgen.mpg.de
authors.forename: A
authors.initials: A
authors.lastname: Guerasimova
authors.forename: L
authors.initials: L
authors.lastname: Nyarsik
authors.forename: I
authors.initials: I
authors.lastname: Girnus
authors.forename: M
authors.initials: M
authors.lastname: Steinfath
authors.forename: W
authors.initials: W
authors.lastname: Wruck
authors.forename: H
authors.initials: H
authors.lastname: Griffiths
authors.forename: R
authors.initials: R
authors.lastname: Herwig
authors.forename: C
authors.initials: C
authors.lastname: Wierling
authors.forename: J
authors.initials: J
authors.lastname: O'Brien
authors.forename: H
authors.initials: H
authors.lastname: Eickhoff
authors.forename: H
authors.initials: H
authors.lastname: Lehrach
authors.forename: U
authors.initials: U
authors.lastname: Radelof
mesh_terms: D001322:Autoanalysis; D016172:DNA Fingerprinting; D015342:DNA Probes; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry*; D020135:Peptide Nucleic Acids; D010799:Phycoerythrin; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D015342:DNA Probes; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; D010799:Phycoerythrin