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 and Helen Donis-Keller |
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| Institution: | 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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