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Nucleic Acids Research 2005 33(1):e4; doi:10.1093/nar/gni007
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Published online 7 January 2005

© 2005, the authors Nucleic Acids Research, Vol. 33 No. 1 © Oxford University Press 2005; all rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use permissions, please contact journals.permissions{at}oupjournals.org.

Methods Online

Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions

Kae Sato, Kazuo Hosokawa and Mizuo Maeda*

Bioengineering Laboratory, RIKEN (The Institute of Physical and Chemical Research) Hirosawa 2-1, Wako, Saitama 351-0198, Japan

*To whom correspondence should be addressed. Tel: +81 48 467 9312; Fax: +81 48 462 4658; Email: mizuo{at}riken.go.jp

Received October 6, 2004. Revised November 28, 2004. Accepted December 13, 2004.

Aggregation of DNA-modified gold nanoparticles in a non-cross-linking configuration has extraordinary selectivity against terminal mismatch of the surface-bound duplex. In this paper, we demonstrate the utility of this selectivity for detection of single-base substitutions. The samples were prepared through standard protocols: DNA extraction, PCR amplification and single-base primer extension. Oligonucleotide-modified nanoparticles correctly responded to the unpurified products from the primer extension: aggregation for the full match and dispersion for all the mismatches. Applicability of this method to genomic DNA was tested with five human tumor cell lines, and verified by conventional technologies: mass spectrometry and direct sequencing. Unlike the existing methods for single-base substitution analysis, this method does not need specialized equipments, and opens up a new possibility of point-of-care diagnosis for single-nucleotide polymorphisms.


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