Spatially Defined Oligonucleotide Arrays. Technical Report for Phase II

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The goal of the Human Genome Project is to sequence all 3 billion base pairs of the human genome. Progress in this has been rapid; GenBank{reg_sign} finished 1994 with 286 million bases of sequence and grew by 2470 in the first quarter of 1995. The challenge to the scientific community is to understand the biological relevance of this genetic information. In most cases the sequence being generated for any single region of the genome represents the genotype of a single individual. A complete understanding of the function of specific genes and other regions of the genome and their role in ... continued below

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7 p.

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Creator: Unknown. June 15, 2000.

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The goal of the Human Genome Project is to sequence all 3 billion base pairs of the human genome. Progress in this has been rapid; GenBank{reg_sign} finished 1994 with 286 million bases of sequence and grew by 2470 in the first quarter of 1995. The challenge to the scientific community is to understand the biological relevance of this genetic information. In most cases the sequence being generated for any single region of the genome represents the genotype of a single individual. A complete understanding of the function of specific genes and other regions of the genome and their role in human disease and development will only become apparent when the sequence of many more individuals is known. Access to genetic information is ultimately limited by the ability to screen DNA sequence. Although the pioneering sequencing methods of Sanger et al. (15) and Maxam and Gilbert (11) have become standard in virtually all molecular biology laboratories, the basic protocols remain largely unchanged. The throughput of this sequencing technology is now becoming the rate-limiting step in both large-scale sequencing projects such as the Human Genome Project and the subsequent efforts to understand genetic diversity. This has inspired the development of advanced DNA sequencing technologies (9), Incremental improvements to Sanger sequencing have been made in DNA labeling and detection. High-speed electrophoresis methods using ultrathin gels or capillary arrays are now being more widely employed. However, these methods are throughput-limited by their sequential nature and the speed and resolution of separations. This limitation will become more pronounced as the need to rapidly screen newly discovered genes for biologically relevant polymorphisms increases. An alternative to gel-based sequencing is to use high-density oligonucleotide probe arrays. Oligonucleotide probe arrays display specific oligonucleotide probes at precise locations in a high density, information-rich format (5,4,1 2). The hybridization pattern of a fluorescently labeled nucleic acid target is used to gain primary structure information of the target. This format can be applied to a broad range of nucleic acid sequence analysis problems including pathogen identification, polymorphism detection, human identification, mRNA expression monitoring and de novo sequencing. In this review, we briefly describe the method of light-directed chemid synthesis to create high-density arrays of oligonucleotide probes, the method of fluorescently labeling target nucleic acids for hybridization to the probe arrays, the detection of hybridized targets by epi-fluorescence confocal scanning and the data analysis procedures used to interpret the hybridization signals. To illustrate the use of specific high-density oligonucleotide probe arrays, we describe their application to screening the reverse transcriptase (rt) and protease (pro) genes of HIV-I for polymorphisms and drug-resistance conferring mutations.

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7 p.

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OSTI as DE00761463

Medium: P; Size: 7 pages

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  • Other Information: PBD: 15 Jun 2000

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  • Report No.: None
  • Grant Number: FG03-92ER81275
  • DOI: 10.2172/761463 | External Link
  • Office of Scientific & Technical Information Report Number: 761463
  • Archival Resource Key: ark:/67531/metadc723135

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Creation Date

  • June 15, 2000

Added to The UNT Digital Library

  • Sept. 29, 2015, 5:31 a.m.

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  • April 11, 2017, 3:45 p.m.

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Spatially Defined Oligonucleotide Arrays. Technical Report for Phase II, report, June 15, 2000; United States. (digital.library.unt.edu/ark:/67531/metadc723135/: accessed August 17, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.