UBC Theses and Dissertations
Mapping of D8S136 to the short arm of human chromosome 8 Mitchell, Heather Katherine
Cosmid clones containing NotI sites and (GT)n repeats were randomly isolated from the flow-sorted human chromosome 8 library, LA08NC01, by hybridization with nick-translated poly(dC-dA)- (dG-dT) and subsequent EcoRI/Notl and NotI digestion of GT-hybridizing clones. An anonymous DNA sequence, D8S136, containing three (GT)n repeats and two NotI sites was isolated. Three sequence tagged sites (STSs), unique sequences identifying D8S136, were generated and the physical distances separating the sites were determined. Two of the (GT)n repeats within this segment were shown to be highly polymorphic in DNA from members of the Centre d'Etude Polymorphisme Humain (CEPH) panel of 40 reference families. These polymorphisms were designated D8S136P1 and D8S136P2. Southern blot analysis and amplification of DNA from somatic cell hybrid lines using the polymerase chain reaction (PCR) mapped D8S136 to 8p, interval 8p21^cen and linkage analysis localized this marker to an approximately 9.9 centimorgan (cM) region between flanking marker loci D8S133 and D8S5. Analysis of allelic association between D8S136P1 and D8S136P2 indicated minimal disequilibrium. The disequilibrium did not appear to measurably decrease the amount of information gained by typing individuals for both D8S136P1 and D8S136P2. The PIC values for D8S136P1 and D8S136P2 were 0.82 and 0.69, respectively. The PIC of the haplotype was 0.95. D8S136 is an extremely polymorphic locus which will be useful for physical and linkage mapping studies.
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