DNA microarrays based on noncovalent oligonucleotide attachment and hybridization in two dimensions |
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Authors: | Belosludtsev Y Iverson B Lemeshko S Eggers R Wiese R Lee S Powdrill T Hogan M |
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Institution: | Genometrix Inc., 3608 Research Forest Drive, Suite B7, The Woodlands, Texas 77381, USA. yurib@genometrix.com |
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Abstract: | Short oligonucleotide probes have been linked to a solid support by simple electrostatic adsorption onto a positively charged surface film. Attachment was obtained by microfluidic application of unmodified oligonucleotides in distilled water onto amino-silanized glass. It has been demonstrated that an extremely stable monolayer of oligonucleotide is obtained by this method, at a density of about 10(11) molecules/mm(2), which approaches the limit expected for a two-dimensional closest-packed array. Application of oligonucleotide by adsorption is followed by capping with acetic anhydride in the vapor phase, and then capping with succinic anhydride in solution to form a surface with weak negative charge. The capping method has been successfully employed for microarray fabrication and for the analysis of single nucleotide polymorphisms in the k-ras gene. The data reveal that, subsequent to capping, the adsorptive association of oligonucleotide to the surface yields a probe layer which is capable of single nucleotide base mismatch discrimination and high apparent binding affinity. |
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