ABSTRACT
High-density DNA probe arrays provide a massively parallel approach to nucleic acid sequence analysis that is transforming gene-based biomedical research and diagnostics. Light-directed combinatorial oligonucleotide synthesis has enabled the large-scale production of GeneChip probe arrays which contain several hundred of thousand oligonucleotide sequences on glass "chips" about one cm2 in size. Due to their very high information content, GeneChip probe arrays are finding widespread use in the hybridization-based detection and analysis of mutations and polymorphisms ("genotyping"), and in a wide range of gene expression studies. The manufacturing process integrates solid-phase photochemical oligonucleotide synthesis with lithographic techniques adapted from the microelectronics industry. The present-generation methodology employs MeNPOC photo-activatable nucleoside monomers with proximity photolithography, and is currently capable of printing individual 10 microns 2 probe features at a density of 10(6) probes/cm2.
Subject(s)
Oligonucleotide Array Sequence Analysis , Oligonucleotide Probes/chemical synthesis , DNA Probes/chemical synthesis , Fluorescein/chemistry , PhotochemistrySubject(s)
2-Aminopurine/analogs & derivatives , Oligonucleotide Array Sequence Analysis/methods , 2-Aminopurine/chemistry , 2-Aminopurine/radiation effects , Amides/chemistry , Amides/radiation effects , Combinatorial Chemistry Techniques/instrumentation , Membrane Glycoproteins , Oligonucleotides/chemical synthesis , Oligonucleotides/radiation effects , Phosphoric Acids/chemistry , Phosphoric Acids/radiation effects , Photolysis , Purine Nucleosides/chemistry , Purine Nucleosides/radiation effectsABSTRACT
DNA probe arrays were synthesized with analogs of 2,6-diaminopurine and 2'-O-methyl-thymidine in place of A and T. AT-rich GeneChip test arrays containing 14-mer or 20-mer analog probes improved hybridization to fluorescently-labeled RNA sequences under stringent conditions.
