Genetic analyses of restriction fragment length polymorphisms using high molecular weight DNA from sperm or lymphocytes embedded in agarose |
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| Affiliation: | 1. Department of Pathology, New York University School of Medicine, Alexandria ERSP, 450 East 29th Street, New York, NY 10016, USA;2. Department of Pathology, The Center for Cognitive Neurology, New York University School of Medicine, Alexandria ERSP, 450 East 29th Street, New York, NY 10016, USA;3. Department of Neurology, The Center for Cognitive Neurology, New York University School of Medicine, Alexandria ERSP, 450 East 29th Street, New York, NY 10016, USA;4. Department of Psychiatry, The Center for Cognitive Neurology, New York University School of Medicine, Alexandria ERSP, 450 East 29th Street, New York, NY 10016, USA;1. Methodology Center, Edna Bennett Pierce Prevention Research Center, Pennsylvania State University, 404 Health and Human Development Building, University Park, PA 16802, United States;2. Center on Young Adult Health and Development, Department of Behavioral and Community Health, University of Maryland School of Public Health, 4200 Valley Dr, #1234, College Park, MD 20742, United States;3. Department of Behavioral and Community Health, University of Maryland School of Public Health, 4200 Valley Dr, #1234, College Park, MD 20742, United States;1. Shaanxi Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China;2. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, Guangxi, China;1. Rutgers University, Piscataway, NJ, USA;1. Department of Family Medicine and Community Health, University of Wisconsin, 1100 Delaplaine Ct., Madison, WI 53715, USA;2. Department of Medicine, Division of Endocrinology, Diabetes & Metabolism, University of Wisconsin, Medical Foundation Centennial Building, Room 4170, 1685 Highland Avenue, Madison, WI 53705, USA |
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| Abstract: | A simple and efficient method for determining restriction fragment length polymorphism types on large numbers of individuals using small samples of peripheral blood or sperm cells is described. Whole cells embedded in low gelling/melting temperature agarose were treated with a series of enzyme, detergent, and washing steps to release high molecular weight DNA that was then digested with standard restriction enzymes such as EcoRI and PstI, electrophoresed, blotted, and probed as in normal Southern analyses. The technique should be readily adaptable to any application requiring DNA from small numbers of cells for Southern analyses or pulsed field gel electrophoresis. |
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