High Throughput Fingerprint Analysis of Large-Insert Clones
- Marco A. Marra1,5,
- Tamara A. Kucaba1,
- Nicole L. Dietrich1,
- Eric D. Green2,
- Buddy Brownstein4,
- Richard K. Wilson1,3,
- Ken M. McDonald1,3,
- LaDeana W. Hillier1,3,
- John D. McPherson1,3, and
- Robert H. Waterston1,3
- 1Washington University School of Medicine, Genome Sequencing Center, St. Louis, Missouri 63108; 2National Human Genome Research Institute (NHGRI), National Institutes of Health, Bethesda, Maryland 20892; 3Department of Genetics and 4Center for Genetics in Medicine, Washington University School of Medicine, St. Louis, Missouri 63108
Abstract
As part of the Human Genome Project, the Washington University Genome Sequencing Center has commenced systematic sequencing of human chromsome 7. To organize and supply the effort, we have undertaken the construction of sequence-ready physical maps for defined chromosomal intervals. Map construction is a serial process composed of three main activities. First, candidate STS-positive large-insert PAC and BAC clones are identified. Next, these candidate clones are subjected to fingerprint analysis. Finally, the fingerprint data are used to assemble sequence-ready maps. The fingerprinting method we have devised is key to the success of the overall approach. We present here the details of the method and show that the fingerprints are of sufficient quality to permit the construction of megabase-size contigs in defined regions of the human genome. We anticipate that the high throughput and precision characteristic of our fingerprinting method will make it of general utility.
Footnotes
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↵5 Corresponding author.
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This paper is dedicated to the memory of Amerigo Marra, 1929–1997.
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E-MAIL mmarra{at}watson.wustl.edu; FAX (314) 286-1810.
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- Received May 5, 1997.
- Accepted September 12, 1997.
- Cold Spring Harbor Laboratory Press
