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Combinatorial Chemistry & High Throughput Screening


ISSN (Print): 1386-2073
ISSN (Online): 1875-5402

LNA-Modified Oligodeoxynucleotide Hybridization with DNA Microarrays Printed on Nanoporous Membrane Slides

Author(s): Hans Lehrach, Regine Schwartz, Matthias Lange, Lajos Nyarsik, Michal Janitz, Anna Guerasimova and Jian-Ping Liu

Volume 9, Issue 8, 2006

Page: [591 - 597] Pages: 7

DOI: 10.2174/138620706778249730

Price: $65


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 ∼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.

Keywords: LNA, oligodeoxynucleotide, hybridization, microarray, nanoporous membrane, re-sequencing

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