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

Nucleic Acids Research
F CattaruzzaEmanuele Chirivino

Abstract

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 parall...Continue Reading

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Citations

Mar 14, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Siyuan ChenLloyd M Smith
Oct 31, 2006·Omics : a Journal of Integrative Biology·Soon Jin OhJoon Won Park
Jul 8, 2008·Analytica Chimica Acta·Sukdeb PalJoon Myong Song
Jul 30, 2011·ACS Applied Materials & Interfaces·Rebecca A ShircliffHoward M Branz
Sep 21, 2007·Journal of the American Chemical Society·Robin L Hayes, Mark E Tuckerman

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Methods Mentioned

BETA
biosensors
fluorescence microscopy
Atomic Force Microscopy
AFM
amidation
biosensor

Software Mentioned

Oligonucletide Properties Calculator

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