Strategies in the preparation of DNA oligonucleotide arrays for diagnostic applications.
This report emphasizes the interfacial chemistry that is required to ensure proper attachment of oligonucleotides onto the surface of microarrays. For example, strategies for the covalent attachment of pre-synthesized oligonucleotides to glass slides, gold films, polyacrylamide gel pads, polypyrrole films, and optical fibers are surveyed in an attempt to better define the parameters for optimal formation and detection of DNA hybrids. These parameters include among others, the nature and length of the linkers attaching oligonucleotides to the arrays, and the surface density of oligonucleotides required for unhindered hybridization with DNA targets. Sensitive detection methods such as the use of light-scattering techniques, molecular beacons, surface plasmon resonance, attenuated total internal reflection-FTIR, and the evanescent field excitation of fluorescence from surface-bound fluorophores have been developed to study the kinetics and specificity of hybridization events. Finally, the synthesis of oligonucleotides directly on glass surfaces and polypropylene sheets has been investigated to enable DNA sequencing by hybridization and achieve oligonucleotide densities of ca. 10(6) sequences per cm(2) on DNA chips.
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Division of Therapeutic Proteins, Center for Biologics Evaluation and Research, Food and Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, USA.
D015342:DNA Probes / Q000737:chemistry*; D005336:Fiber Optic Technology; D015870:Gene Expression; D005898:Glass / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D007103:Immobilization; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D009841:Oligonucleotides / Q000138:chemical synthesis*; D055100:Optical Fibers; D011126:Polypropylenes / Q000737:chemistry*; D013499:Surface Properties
D015342:DNA Probes; D009841:Oligonucleotides; D011126:Polypropylenes; D006046:Gold