High density synthetic oligonucleotide arrays

Experimental genomics involves taking advantage of sequence information to investigate and understand the workings of genes, cells and organisms. We have developed an approach in which sequence information is used directly to design high–density, two–dimensional arrays of synthetic oligonucleotides. The GeneChip® probe arrays are made using spatially patterned, light–directed combinatorial chemical synthesis, and contain up to hundreds of thousands of different oligonucleotides on a small glass surface. The arrays have been designed and used for quantitative and highly parallel measurements of gene expression, to discover polymorphic loci and to detect the presence of thousands of alternative alleles. Here, we describe the fabrication of the arrays, their design and some specific applications to high–throughput genetic and cellular analysis.

[1]  S. P. Fodor,et al.  Light-directed, spatially addressable parallel chemical synthesis. , 1991, Science.

[2]  E. Southern,et al.  Analyzing and comparing nucleic acid sequences by hybridization to arrays of oligonucleotides: evaluation using experimental models. , 1992, Genomics.

[3]  S. P. Fodor,et al.  Multiplexed biochemical assays with biological chips , 1993, Nature.

[4]  S. P. Fodor,et al.  Light-generated oligonucleotide arrays for rapid DNA sequence analysis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Ronald W. Davis,et al.  Quantitative phenotypic analysis of yeast deletion mutants using a highly parallel molecular bar–coding strategy , 1996, Nature Genetics.

[6]  G. Mcgall,et al.  Light-directed synthesis of high-density oligonucleotide arrays using semiconductor photoresists. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[7]  James L. Winkler,et al.  Accessing Genetic Information with High-Density DNA Arrays , 1996, Science.

[8]  N. Shen,et al.  Extensive polymorphisms observed in HIV–1 clade B protease gene using high–density oligonucleotide arrays , 1996, Nature Medicine.

[9]  D. Lockhart,et al.  Expression monitoring by hybridization to high-density oligonucleotide arrays , 1996, Nature Biotechnology.

[10]  R. Sapolsky,et al.  Mapping genomic library clones using oligonucleotide arrays. , 1996, Genomics.

[11]  S. P. Fodor DNA SEQUENCING: Massively Parallel Genomics , 1997, Science.

[12]  L. Wodicka,et al.  Genome-wide expression monitoring in Saccharomyces cerevisiae , 1997, Nature Biotechnology.

[13]  Nam Quoc Ngo,et al.  The Efficiency of Light-Directed Synthesis of DNA Arrays on Glass Substrates , 1997 .

[14]  Antoine de Saizieu,et al.  Bacterial transcript imaging by hybridization of total RNA to oligonucleotide arrays , 1998, Nature Biotechnology.

[15]  T. Gingeras,et al.  Cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S H Kim,et al.  Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors. , 1998, Science.

[17]  D. Richman,et al.  Comparative performance of high-density oligonucleotide sequencing and dideoxynucleotide sequencing of HIV type 1 pol from clinical samples. , 1998, AIDS research and human retroviruses.

[18]  Ronald W. Davis,et al.  A genome-wide transcriptional analysis of the mitotic cell cycle. , 1998, Molecular cell.

[19]  C. Nusbaum,et al.  Large-scale identification, mapping, and genotyping of single-nucleotide polymorphisms in the human genome. , 1998, Science.

[20]  D. Lockhart,et al.  Deciphering Molecular Circuitry Using High-Density DNA Arrays , 1998 .

[21]  A. Berno,et al.  Simultaneous genotyping and species identification using hybridization pattern recognition analysis of generic Mycobacterium DNA arrays. , 1998, Genome research.

[22]  L Wodicka,et al.  Parallel analysis of genetic selections using whole genome oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[23]  K. Gunderson,et al.  Mutation detection by ligation to complete n-mer DNA arrays. , 1998, Genome research.

[24]  M. Pirrung,et al.  Proofing of Photolithographic DNA Synthesis with 3‘,5‘-Dimethoxybenzoinyloxycarbonyl-Protected Deoxynucleoside Phosphoramidites , 1998 .

[25]  Daniel R. Richards,et al.  Direct allelic variation scanning of the yeast genome. , 1998, Science.

[26]  M. Bittner,et al.  Expression profiling using cDNA microarrays , 1999, Nature Genetics.

[27]  D. Bowtell,et al.  Options available—from start to finish—for obtaining expression data by microarray , 1999, Nature Genetics.

[28]  A. Troesch,et al.  Mycobacterium Species Identification and Rifampin Resistance Testing with High-Density DNA Probe Arrays , 1999, Journal of Clinical Microbiology.

[29]  J. Hacia Resequencing and mutational analysis using oligonucleotide microarrays , 1999, Nature Genetics.

[30]  A. Chakravarti Population genetics—making sense out of sequence , 1999, Nature Genetics.

[31]  M. Morley,et al.  Making and reading microarrays , 1999, Nature Genetics.