Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm: experimental design considerations.
The 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.
Boehnke
M
M
Department of Biostatistics, School of Public Health, University of Michigan, Ann Arbor 48109.
Arnheim
N
N
Li
H
H
Collins
F S
FS
D000483:Alleles; D002874:Chromosome Mapping; D002877:Chromosomes, Human; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D005787:Gene Frequency; D006801:Humans; D008297:Male; D015233:Models, Statistical; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D011336:Probability; D013094:Spermatozoa / Q000032:analysis*
D004259:DNA-Directed DNA Polymerase