A DNA microarray system for analyzing complex DNA samples using two-color fluorescent probe hybridization.

Genome Research - Tập 6 Số 7 - Trang 639-645 - 1996
Dari Shalon1, Steven J. Smith1, Patrick O. Brown1
11Howard Hughes Medical Institute and Departments of 2Biochemistry and 3Molecular and Cellular Physiology, Stanford University, Stanford, California 94305

Tóm tắt

Detecting and determining the relative abundance of diverse individual sequences in complex DNA samples is a recurring experimental challenge in analyzing genomes. We describe a general experimental approach to this problem, using microscopic arrays of DNA fragments on glass substrates for differential hybridization analysis of fluorescently labeled DNA samples. To test the system, 864 physically mapped lambda clones of yeast genomic DNA, together representing >75% of the yeast genome, were arranged into 1.8-cm x 1.8-cm arrays, each containing a total of 1744 elements. The microarrays were characterized by simultaneous hybridization of two different sets of isolated yeast chromosomes labeled with two different fluorophores. A laser fluorescent scanner was used to detect the hybridization signals from the two fluorophores. The results demonstrate the utility of DNA microarrays in the analysis of complex DNA samples. This system should find numerous applications in genome-wide genetic mapping, physical mapping, and gene expression studies.

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Tài liệu tham khảo

1991, New techniques for physical mapping of the human genome., FASEB J., 5, 28, 10.1096/fasebj.5.1.1846833

10.1016/0888-7543(92)90057-Y

10.1126/science.3538420

10.1126/science.8503011

10.1093/nar/21.5.1321

10.1126/science.270.5244.1945

10.1126/science.1359641

10.1093/nar/20.7.1675

1995, Genomic mismatch scanning: A new approach to genetic linkage mapping,, Nature Genet., 4, 11

10.1073/pnas.91.11.5022

1993, Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs., Genetics, 134, 81, 10.1093/genetics/134.1.81

Ross, M.T., J.D. Hoheisel, A.P. Monaco, Z. Larin, G. Zehetner, and H. Lehrach. 1992. High density gridded YAC filters: Their potential as genome mapping tools. In Techniques for the analysis of complex genomes (ed. Rakesh Anand), pp. 137–153. Academic Press, London, UK.

10.1126/science.270.5235.467

Shalon, D., 1995. “DNA micro arrays: A new tool for genetic analysis.” Ph.D. thesis, Stanford University, Stanford, CA.

10.1038/367489a0

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Genome Research

Tập 6 Số 7

639-645

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