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Nucleic Acids Research 2006 34(4):e32; doi:10.1093/nar/gnj034
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Published online 28 February 2006

© The Author 2006. Published by Oxford University Press. 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, please contact journals.permissions{at}oxfordjournals.org

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Controlled loading of oligodeoxyribonucleotide monolayers onto unoxidized crystalline silicon; fluorescence-based determination of the surface coverage and of the hybridization efficiency; parallel imaging of the process by Atomic Force Microscopy

Fabrizio Cattaruzza1, Antonio Cricenti1, Alberto Flamini1,*, Marco Girasole1, Giovanni Longo1, Tommaso Prosperi1, Giuseppina Andreano2, Luciano Cellai2 and Emanuele Chirivino2

1Istituto di Struttura della Materia, CNR Via Salaria Km 29,300, 00016 Monterotondo Stazione, Rome, Italy 2Istituto di Cristallografia, CNR Via Salaria Km 29,300, 00016 Monterotondo Stazione, Rome, Italy

*To whom correspondence should be addressed. Tel: +39 06 90672318; Fax: +39 06 90672316; Email: alberto.flamini{at}ism.cnr.it

Received November 10, 2005. Revised December 23, 2005. Accepted February 4, 2006.

Unoxidized crystalline silicon, characterized by high purity, high homogeneity, sturdiness and an atomically flat surface, offers many advantages for the construction of electronic miniaturized biosensor arrays upon attachment of biomolecules (DNA, proteins or small organic compounds). This allows to study the incidence of molecular interactions through the simultaneous analysis, within a single experiment, of a number of samples containing small quantities of potential targets, in the presence of thousands of variables. A simple, accurate and robust methodology was established and is here presented, for the assembling of DNA sensors on the unoxidized, crystalline Si(100) surface, by loading controlled amounts of a monolayer DNA-probe through a two-step procedure. At first a monolayer of a spacer molecule, such as 10-undecynoic acid, was deposited, under optimized conditions, via controlled cathodic electrografting, then a synthetic DNA-probe was anchored to it, through amidation in aqueous solution. The surface coverage of several DNA-probes and the control of their efficiency in recognizing a complementary target-DNA upon hybridization were evaluated by fluorescence measurements. The whole process was also monitored in parallel by Atomic Force Microscopy (AFM).

Correspondence may also be addressed to Luciano Cellai. Tel: +39 06 90672613; Fax: +39 06 90672630; Email: luciano.cellai{at}ic.cnr.it


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