Keywords
DNA sequencing, Forensic, Pyrosequencing, SNP, Sequencing, Single nucleotide polymorphism, Y chromosome
Abstract
The potential value of the Y chromosome for forensic applications has been recognized for some time with the current work dedicated to Short Tandem Repeat analysis and Single Nucleotide Polymorphism (SNP) discovery. This study examined the ability of two different SNP analysis methods to determine if they could be utilized in forensic applications and ultimately be developed into an established system for Y chromosome SNP analysis. This study examined two principle SNP analysis systems: single base extension and Pyrosequencing. Pyrosequencing was determined to be superior to single base extension, due to the wealth of information provided with sequencing and the flexibility of designing primers for analysis. Using Pyrosequencing, 50 Y chromosome loci were examined and the minimum loci required for maximum diversity for the development of a Y chromosome SNP analysis system were chosen. Thirteen loci were selected based on their ability to discriminate 60 different individuals from three different racial groups into 15 different haplogroups. The Y chromosome SNP analysis system developed utilized nested PCR for the amplification of all 13 loci. Then they were sequenced as groups, ranging from one to three loci, in a single reaction. The Y chromosome SNP analysis system developed here has the potential for forensic application since it has shown to be successful in the analysis of blood, buccal swabs, semen, and saliva, works with as little as 5 pg of starting DNA material, and will amplify only male DNA in the presence of male/female mixtures in which the female portion of the sample overwhelmed the male portion 30,000 to 1.
Notes
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Graduation Date
2004
Semester
Summer
Advisor
Ballantyne, Jack
Degree
Master of Science (M.S.)
College
College of Arts and Sciences
Department
Chemistry
Degree Program
Chemistry
Format
application/pdf
Identifier
CFE0000071
URL
http://purl.fcla.edu/fcla/etd/CFE0000071
Language
English
Release Date
August 2004
Length of Campus-only Access
None
Access Status
Masters Thesis (Open Access)
Subjects
Arts and Sciences -- Dissertations, Academic; Dissertations, Academic -- Arts and Sciences
STARS Citation
Fletcher, Jeremy Charles, "The Use Of Pyrosequencing For The Analysis Of Y Chromosome Single Nucleotide Polymorphisms" (2004). Electronic Theses and Dissertations. 27.
https://stars.library.ucf.edu/etd/27