The Polymerase Chain Reaction will be used to analyze DNA sequences in single human sperm. The ability to study large numbers of meiotic products from single individuals will allow the construction of human genetic maps at a resolution which far exceeds that possible by using conventional pedigree analysis. Sperm will be isolated using flow cytometry. The efficiency of sperm lysis and PCR amplification will be maximized under conditions which minimize contamination. The immediate goal is to be able to routinely achieve a level of resolution for genetic mapping of less than 0.1 cM (0.001 recombination fraction). Accomplishment of this goal will allow three point crosses to be used to order very closely linked markers. The relationship between physical distance and recombination frequency could also be investigated. The development of this single sperm technology may have a significant impact on mapping the human genome.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
8R01HG000328-02
Application #
3333439
Study Section
Special Emphasis Panel (SRC)
Project Start
1989-07-01
Project End
1992-06-30
Budget Start
1990-07-01
Budget End
1991-06-30
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Southern California
Department
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
Zhang, L; Cui, X; Schmitt, K et al. (1992) Whole genome amplification from a single cell: implications for genetic analysis. Proc Natl Acad Sci U S A 89:5847-51
Lewin, H A; Schmitt, K; Hubert, R et al. (1992) Close linkage between bovine prolactin and BoLA-DRB3 genes: genetic mapping in cattle by single sperm typing. Genomics 13:44-8
Cui, X; Gerwin, J; Navidi, W et al. (1992) Gene-centromere linkage mapping by PCR analysis of individual oocytes. Genomics 13:713-7
Navidi, W; Arnheim, N; Waterman, M S (1992) A multiple-tubes approach for accurate genotyping of very small DNA samples by using PCR: statistical considerations. Am J Hum Genet 50:347-59
Hubert, R; Weber, J L; Schmitt, K et al. (1992) A new source of polymorphic DNA markers for sperm typing: analysis of microsatellite repeats in single cells. Am J Hum Genet 51:985-91