Validation of Applied Biosystems 3130xl Genetic Analyzer for Human mtDNA: An Evaluation of Three Amplification Series

dc.contributor.advisorJoseph Warren
dc.contributor.committeeMemberJohn Planz
dc.contributor.committeeMemberArthur Eisenberg
dc.creatorDavid, Jamalia Junelle
dc.description.abstractThe introduction of a new instrument into an accredited laboratory requires a documented internal validation. Validations typically include sensitivity, precision, and mixture studies. These tests assess the reliability and efficiency of the instrument and allow for interpretation guidelines to be established. This project consisted of a validation of Applied Biosystems’ 3130xl Genetic Analyzer and an evaluation of three mitochondrial DNA amplification primer sets for the control region. The validation was designed to evaluate the efficacy, robustness, and working limitations of the 3130xl instrument by performing a sensitivity study. A sensitivity study was performed using DNA sample dilutions, which were quantified using ABIs Quantifiler system to measure the amount of total nuclear DNA content in the samples. The samples were amplified on the GeneAmp PCR System 9700 in triplicate to evaluate stochastic activity. Three different primer sets were utilized which allow for the amplification of different regions of the human mitochondrial genome control region. After amplification, the quality and quantity of the DNA in all the samples was assessed using the Agilent 2100 BioAnalyzer, and subsequent sequence analysis was performed on the 3130xl Genetic Analyzer. Preliminary work has begun on a mixture study, but due to lack of time and reagents, this study was not completed and will have to be performed at a later date. All sequence data from the sensitivity study was evaluated using Sequencher version 4.1.4Fb19
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.subjectaccredited laboratory
dc.subjectinternal validation
dc.subjectApplied Biosystems’ 3130xl Genetic Analyzer
dc.subjectDNA sample dilutions
dc.subjectGeneAmp PCR System 9700
dc.subjectAgilent 2100 BioAnalyzer
dc.subjectsequence data
dc.subjectstochastic activity
dc.subjecthuman mitochondrial genome control
dc.titleValidation of Applied Biosystems 3130xl Genetic Analyzer for Human mtDNA: An Evaluation of Three Amplification Series
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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