The goal of the proposed research is to develop """"""""framework"""""""" maps consisting of highly informative """"""""index"""""""" markers (heterozygosities greater than or equal to 0.70) spaced at 10 - 15 cM intervals for human chromosomes 5 8, and 16. Based on the estimated lengths of these chromosomes, a total of 80 markers will be needed: 30 for chromosomes 5 and 8 and 20 for chromosome 16. To date, 10, 6, and 7 markers on chromosomes 5, 8, and 16, respectively, are sufficiently informative and spaced at appropriate intervals to qualify as index markers. Due to the high resolution of current multipoint maps for chromosomes 5 and 16, the existing loci that qualify as index markers will be supplemented by expanding the informativeness of markers at appropriate intervals on the multipoint maps by searching for polymorphism in the poly A tract of Alu elements and loci with short tandem repeats (STRs). Sub-cloning will be performed on candidate markers and PCR amenable methods of genotyping will be developed. Those loci that have heterozygosities greater than or equal to 0.70 (based on 10 CEPH parents) will be genotyped on the current set of 40 CEPH families by PCR multiplexing technology and incorporated into the genetic map using CRI-MAP. Markers in critical regions that do not reveal additional polymorphisms will be expanded in genomic YAC or cosmid libraries. If necessary, new markers that reveal Alu and STR variation will be isolated from chromosome specific libraries. It is anticipated that this effort will be minimal. Since the current multipoint map for chromosome 8 is not of sufficient resolution to facilitate the choice of candidate markers for a search of additional polymorphisms, the existing markers on this chromosome that exhibit heterozygosities greater than or equal to 0.70 will be supplemented by the isolation, from chromosome 8-specific libraries subcloned into M13, of new markers that reveal variation in Alu elements and STRs. A hybrid panel which divides the chromosome into 10 intervals, will be utilized to expedite the choice of candidate loci. Polymorphic loci will be selected and genotypic data collected and analyzed as described above. The framework maps will be considered complete when telomere probes are incorporated and when the informativeness and spacing of the collected markers meet the criteria specified by this RFA.
| Zheng, C J; Ma, N S; Dorman, T E et al. (1994) Development of 124 sequence-tagged sites and cytogenetic localization of 217 cosmids for human chromosome 10. Genomics 22:55-67 |
