Date of Award

5-1-2015

Degree Type

Thesis

Degree Name

Master of Science

Field of Study

Forensic Genetics

Department

Graduate School of Biomedical Sciences

First Advisor

Arthur J. Eisenberg

Second Advisor

Rhonda Roby

Third Advisor

Dixie L. Peters

Abstract

Evaluation of the control region of the mitochondrial genome is a common practice for forensic casework and research purposes. Since no kit is currently commercially available for the amplification of mitochondrial DNA (mtDNA), its sequencing procedure is time-consuming and laborious. Six steps are generally followed: DNA extraction, quantification and normalization, amplification of two regions (hypervariable regions 1 and 2), cycle sequencing, capillary electrophoresis and data analysis.

This project evaluated a mtDNA direct amplification kit by performing developmental and internal validations. The studies performed included sensitivity, stability, reproducibility, case- type samples, mixtures and accuracy. The mtDNA direct amplification kit successfully amplified reference samples used in each study without the need of extraction and quantification steps. In addition, mtDNA profiles were obtained from the sequenced amplification products. Using the validated direct amplification procedure in the laboratory will improve workflow, decrease operational cost and reduce the possibility of error by minimizing sample handling.

Comments

Alicea-Centeno, Alessandra. Developmental and Internal Validation of a Mitochondrial DNA Direct Amplification Kit for Forensic Reference Samples. Master of Science (Biomedical Sciences, Forensic Genetics). April, 2015, 42 pages, 7 tables, 15 figures, references.