Construction of a Cost Effective Nested-PCR Reaction for Use with the Applied Biosystems AmpFLSTR Identifiler Kit

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dc.contributor.advisorJohn Planz
dc.contributor.committeeMemberJoseph Warren
dc.contributor.committeeMemberArthur Eisenberg
dc.creatorMikeska, Margo M.
dc.date.accessioned2019-08-22T21:29:08Z
dc.date.available2019-08-22T21:29:08Z
dc.date.issued2005-12-01
dc.date.submitted2014-03-18T14:02:27-07:00
dc.description.abstractHuman STR analysis has greatly increased the ability to perform identity testing for many different situations. These situations include, but are not limited to, the identification of individuals involved in violent crimes, establishing paternity, and identification of unknown human remains. The most common type of DNA information currently used for identity testing is the short tandem repeat, or STR. STR testing utilizes the number of repeating units in the DNA to assign an allele. Alleles from several different loci are used to establish a genetic profile. Currently, the United States used a standard of 13 different DNA loci to establish identity. These 13 loci can be typed by using a number of different multiplex kits such as the Applied Biosystems Profiler Plus, Cofiler, and Identifiler Kits [1,2]. The 13 loci were selected based on a number of parameters. Each locus was required to be polymorphyic, and a tetranucleotide repeat. The loci also could not display any linkage between each other and extensive population studies had to be conducted to both verify the absence of linkage and to establish allelic frequencies [1]. The goal of this research was the construction of a more cost effective method of utilizing the Applied Biosystems Identifiler Kit. Across the country there is a large backlog of samples that need to be processed in order to obtain a genetic profile. If these samples could be tested using a more cost effective method, more funding could be directed to other endeavors. Paternity testing, as well as some research endeavors could be conducted at a fraction of the cost, leaving resources for other projects or additional staff. Although it would be inadvisable to use this technique on forensic samples, the implications on paternity and research samples would be positive. This research attempted to design a nested PCR reaction and subsequently dilute the Applied Biosystems Primers in order to reduce the cost. The first step was to design new primers for the first round of PCR, followed by testing of those primers. The new primers then required optimization so that they all worked effectively together. After optimization was accomplished, the Identifiler primers were diluted until loci began dropping out of the genetic profile.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/20.500.12503/29367
dc.language.isoen
dc.provenance.legacyDownloads0
dc.subjectCell and Developmental Biology
dc.subjectCells
dc.subjectComputational Biology
dc.subjectEquipment and Supplies
dc.subjectGenetics
dc.subjectGenetics and Genomics
dc.subjectGenetic Structures
dc.subjectGenomics
dc.subjectHealth Information Technology
dc.subjectInvestigative Techniques
dc.subjectLife Sciences
dc.subjectMedical Cell Biology
dc.subjectMedical Genetics
dc.subjectMedicine and Health Sciences
dc.subjectMolecular Genetics
dc.subjectOther Genetics and Genomics
dc.subjectHuman STR analysis
dc.subjectidentity testing
dc.subjectviolent crimes
dc.subjectpaternity
dc.subjectunknown human remains
dc.subjectDNA
dc.subjectallele
dc.subjectmultiplex kits
dc.subjectpolymorphyic
dc.subjecttetranucleotide
dc.subjectcost effective method
dc.subjectApplied Biosystems Identifiler Kit
dc.subjectnested PCR reaction
dc.subjectcost
dc.subjectoptimization
dc.subjectgenetic profile
dc.titleConstruction of a Cost Effective Nested-PCR Reaction for Use with the Applied Biosystems AmpFLSTR Identifiler Kit
dc.typeProfessional Report
dc.type.materialtext
thesis.degree.departmentGraduate School of Biomedical Sciences
thesis.degree.disciplineCell Biology and Genetics
thesis.degree.grantorUniversity of North Texas Health Science Center at Fort Worth
thesis.degree.nameMaster of Science

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