Identification of more than 500 RFLPs by screening random genomic clones |
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| Authors: | James W. Schumm Robert G. Knowlton Jeffrey C. Braman David F. Barker David Botstein Gita Akots Valerie A. Brown Thomas C. Gravius Cynthia Helms Kathy Hsiao Kenneth Rediker Joan G. Thurston Helen Donis-Keller |
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| Affiliation: | Department of Human Genetics, Collaborative Research, Inc., Bedford, MA 01730. |
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| Abstract: | As part of our genome-mapping effort, we undertook a large-scale screening study to identify RFLPs useful as genetic markers. Some 1,664 single-copy or repeat-containing phage clones from a Charon 4A genomic library were tested for polymorphism against a panel of DNAs, from five unrelated individuals, digested with eight restriction enzymes. Approximately 30% (515) of the clones revealed polymorphism by Southern hybridization; 67 loci detected had PIC values greater than .5. Restriction enzymes MspI, TaqI, and RsaI were most efficient in detecting polymorphism within the 1-20-kb-fragment size range resolved. With only one exception each of the clones detected polymorphism originating from a single locus. |
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