All ETDs from UAB

Advisory Committee Chair

Jason G Linville

Advisory Committee Members

Elizabeth Gardner

M Tino Unlap

Document Type

Thesis

Date of Award

2023

Degree Name by School

Master of Science (MS) College of Arts and Sciences

Abstract

Short Tandem Repeat (STR) typing has been accepted globally as a gold standard for human identity testing. In 1997, the Federal Bureau of Investigation (FBI) Laboratory’s newly developed Combined DNA Index System (CODIS) consisted of 13 core loci to be targeted while DNA typing. In this context, leading biotechnology companies like Promega (Madison WI), Applied Biosystem/Life Technologies (Foster City, CA) etc. offer commercial STR typing kits for crime laboratories. However, these expensive kits are not the most suitable for academic and research laboratories. Students learning the method of STR typing and researchers showing proof of concept do not require identifying 13 loci. Instead, a smaller number of loci can be targeted, reducing the cost for laboratories operating on a low budget. For the past 10 years a custom multiplex STR amplification reaction has been used in the Forensic Science program at the University of Alabama at Birmingham (UAB). This cost-effective reaction has worked well, but supplementing its use with a second reaction would increase the significance of the results and provide a tool for troubleshooting contamination issues in the lab. Additionally, the process for designing and validating the first reaction was not published, so publishing the details of this reaction in a thesis provides a resource for other labs. Hence, in this project an additional reaction targeting 3 new loci was developed and validated. The cost for each 25 μL reaction was reduced to $0.74, much lower than the commercial kits that cost as much as $30 per reaction. To develop the reaction, the three loci D5S818, D16S539 and D18S51 and their corresponding primer sequences were selected. Then, a monoplex reactions for each locus was completed followed by a multiplex reaction. Finally, a validation study (sensitivity study, thermal cycling parameter selection, precision study, accuracy, and concordance study) was performed. The project has resulted in a new, inexpensive, validated multiplex STR amplification reaction that is efficient, and can be used by academicians and researchers in the field of forensic science.

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