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Title: Genetic mapping on human chromosome 9 by analysis of meiotic recombination in single sperm using polymorphic microsatellite markers
Author: Brown, G. M.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 1997
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Flow-stored single sperm were amplified by polymerase chain reaction (PCR) to compare the genetic recombination of 4 individuals using a series of polymorphic microsatellite markers across human chromosome 9q31-q34.3. Firstly, a method was developed to circumvent the problem of marker scarcity in certain areas of the genome. The method enriched a chromosome 9-specific plasmid library for [CA]n microsatellite sequences which could then be investigated for heterozygosity and use as genetic markers. 64% of [CA]n sequences identified after the enrichment procedure were found to contain a tract of at least 12 repeat units; the size at which such tandem repeats are thought to have the potential to be polymorphic. 3 clones were selected and characterised. One clone was found to have an estimated heterozygosity of 82% and was designated D9S749. Sperm typing was performed using a protocol involving whole genome amplification using[N]15-mers and single step multiplex of sets of markers on 2 normal donors for a series of markers spanning 9q31-q34.3. Linkage analysis was performed with CRIMAP, adapted for sperm typing, and the sperm typing program SPERM. The marker order found for 1 donor was cen-D9S109-D9S59-D9S170-D9S154-D9S315-ASS-D9S149-D9S67-ter. This data was in agreement with previously published data but the ordering of D9S170 centromeric to D9S154 had not been previously reported. The same order was found for the marker set of the second donor. Some heterogeneity was found in the % recombination between these 2 individuals with significant differences in recombination frequencies between some markers.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available