title: LNA-modified oligodeoxynucleotide hybridization with DNA microarrays printed on nanoporous membrane slides.
abstract: We report a robust method for the detection of hybridization events using a microarray-based assay on a nanoporous membrane platform. The technique is characterized by a hybridization time of only 1 hour and uses Cy5-labeled, 7-mer oligodeoxynucleotide probes modified with locked nucleic acid (LNA) nucleotides. We show that the volume of the DNA spotted onto a nanomembrane can be reduced to approximately 4 nL with detectable signal intensity. Moreover, the amount of the DNA target could be reduced to 4 fmol. The described approach could dramatically increase the throughput of techniques based on sequencing by hybridization, such as oligofingerprinting, by decreasing the total number of probes that are needed for analysis of large clone sets and reduction of the sample/reagent consumption. The method is particularly advantageous when numerous hybridization-based assays must be performed for characterization of sample sets of 100,000 or more.
authors.affiliation: Department of Vertebrate Genomics, Max-Planck-Institute for Molecular Genetics, D-14195 Berlin, Germany.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Liu
authors.forename: Anna
authors.initials: A
authors.lastname: Guerasimova
authors.forename: Regine
authors.initials: R
authors.lastname: Schwartz
authors.forename: Matthias
authors.initials: M
authors.lastname: Lange
authors.forename: Hans
authors.initials: H
authors.lastname: Lehrach
authors.forename: Lajos
authors.initials: L
authors.lastname: Nyársik
authors.forename: Michal
authors.initials: M
authors.lastname: Janitz
mesh_terms: D008567:Membranes, Artificial; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D015345:Oligonucleotide Probes; D013997:Time Factors
chemical_list: D008567:Membranes, Artificial; D015345:Oligonucleotide Probes