Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm: experimental design considerations.
1989; National Institutes of Health; Volume: 45; Issue: 1 Linguagem: Inglês
Autores
Michael Boehnke, Norman Arnheim, Hongmei Li‐Byarlay, Francis S. Collins,
Tópico(s)Molecular Biology Techniques and Applications
ResumoThe polymerase chain reaction (PCR) makes it possible to rapidly generate a very large number of copies of a specific region of DNA. Application of PCR to individual human sperm cells permits the typing of a large number of independent male meiotic events. If the donor male is heterozygous at three loci, sperm typing using PCR will permit ordering of loci in a manner analogous to classical methods of experimental genetics. Sequential analysis of trios of loci by sperm typing will provide a very accurate means of ordering any number of tightly linked loci. Here, we describe experimental design and sample-size issues raised by the application of sperm typing by PCR for mapping human chromosomes, and we demonstrate that sperm typing will be an efficient method for generating fine-structure human genetic maps.
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