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 ∼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
Combinatorial Chemistry & High Throughput Screening
Title: LNA-Modified Oligodeoxynucleotide Hybridization with DNA Microarrays Printed on Nanoporous Membrane Slides
Volume: 9 Issue: 8
Author(s): Hans Lehrach, Regine Schwartz, Matthias Lange, Lajos Nyarsik, Michal Janitz, Anna Guerasimova and Jian-Ping Liu
Affiliation:
Keywords: LNA, oligodeoxynucleotide, hybridization, microarray, nanoporous membrane, re-sequencing
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 ∼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.
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Cite this article as:
Lehrach Hans, Schwartz Regine, Lange Matthias, Nyarsik Lajos, Janitz Michal, Guerasimova Anna and Liu Jian-Ping, LNA-Modified Oligodeoxynucleotide Hybridization with DNA Microarrays Printed on Nanoporous Membrane Slides, Combinatorial Chemistry & High Throughput Screening 2006; 9 (8) . https://dx.doi.org/10.2174/138620706778249730
DOI https://dx.doi.org/10.2174/138620706778249730 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
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