Analysis of Low Copy Number DNA Using Profiler Plus at Increased Amplification Cycles and Modifications in Sample Injection Parameters

dc.contributor.advisorArthur Eisenberg
dc.contributor.committeeMemberJohn Planz
dc.contributor.committeeMemberJoseph Warren
dc.creatorHynds, Jody Lynn
dc.description.abstractThere are many DNA testing techniques that can be utilized for samples with low quantities of DNA. Mitochondrial DNA testing is designed for successful DNA sequencing of hair shafts, degraded and burned samples. Newly developed SNP (single nucleotide polymorphisms) testing is also designed for the analysis of challenging samples. The increased interest in the analysis of low copy number DNA samples using STR testing is necessitated since the national database CODIS (Combined Data Index System) currently only accepts the DNA profiles analyzed with the 13 core STR loci. CODIS contains DNA profiles of evidence found at crime scenes, convicted offender and missing persons DNA profiles (4). The goal of this project is to develop methodologies to increase the success rate of LCN DNA samples using STR testing. The experimental design for this study involved the amplification of DNA isolated from buccal swabs using the Profiler Plus multiplex kit at two different DNA input quantities: 0/0156ng (15.6pg) and 0.0312ng (31.2pg). Four separate amplifications of these DNA samples were done at: 28, 30, 32 and 34 cycles. The manufacturer’s recommended cycle number for AmpFISTR Profiler Plus is 28 cycles. These samples were analyzed on both the ABI Prism 310 Genetic Analyzer and the ABI Prism 3100 Genetic Analyzer using OCD standard protocols for loading samples. The injection time and voltage were modified for each of the number of PCR cycles. The best combination of cycle number and injection parameters was chosen for the low copy number reproducibility study.
dc.subjectCell and Developmental Biology
dc.subjectComputational Biology
dc.subjectEquipment and Supplies
dc.subjectForensic Science and Technology
dc.subjectGenetics and Genomics
dc.subjectGenetic Structures
dc.subjectHealth Information Technology
dc.subjectInvestigative Techniques
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.subjectOther Genetics and Genomics
dc.subjectDNA testing techniques
dc.subjectmitochondrial DNA testing
dc.subjectdegraded samples
dc.subjectchallenging samples
dc.subjectchallenged samples
dc.subjectlow copy number DNA
dc.subjectSTR testing
dc.subjectbuccal swabs
dc.subjectProfiler Plus multiplex kit
dc.subjectAmpFlSTR Profiler Plus
dc.subjectPCR cycles
dc.titleAnalysis of Low Copy Number DNA Using Profiler Plus at Increased Amplification Cycles and Modifications in Sample Injection Parameters
dc.typeProfessional Report
dc.type.materialtext School of Biomedical Sciences Biology and Genetics of North Texas Health Science Center at Fort Worth of Science


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