Discipline: Biological Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Nena W. Hawkins - University of Mississippi
Co-Author(s): Dr. Murrell Godfrey, University of Mississippi, University, MS; Rachel Boone, University of Mississippi, University, MS; Caroline Spencer, University of Mississippi, University, MS; Ann-Elodie Robert, University of Mississippi, University, MS
There are several different media from which DNA can be extracted. These media can include; blood, saliva, cheek cells, and even bones and teeth. Most DNA and mitochondrial DNA extraction kits are applied to blood or buccal swabbed samples. The aim of this study was to determine if selected extraction kits can be applied to extraction of mitochondrial DNA from teeth. This study will examine two extraction kits, 1) the QIAmp DNA Blood Mini Kit, applied to buccal swabbed samples, used along with a protocol modified by the Dawson-Cruz lab at Virginia Commonwealth University, and 2) A Mitochondrial DNA Polymorphisms in Human Evolution Kit designed by Carolina Biological Supply Company. The first step of this study involved extraction of mitochondrial DNA from the buccal swabs of 5 participates following the protocols of each kit. Once the extraction from buccal samples was complete, amplification and analysis were completed using gel electrophoresis. The successful extraction of the mitochondrial DNA was confirmed through the analysis of the agarose gel. The protocols were then applied to the teeth extractions. Five unknown male teeth and five unknown female teeth were used in this study. These teeth were provided by a local dentist that were extracted within the last 2 years. The teeth were cleaned with a 0.5M EDTA solution and sterilized before the start of each extraction. In order to extract the DNA, a dental drill was used to penetrate the center of the tooth and reach the pulp containing the DNA. Once the sample was ready, the isolation and purification of the mitochondrial DNA (mtDNA) was completed. The mtDNA was then amplified through polymerase chain reaction (PCR). During this process, primers were chosen from the HVI or HVII regions to amplify the mtDNA. After amplification, the PCR products were analyzed through gel electrophoresis to confirm extraction, isolation, and amplification of the mtDNA. DNA analysis was then performed using the ABI 310 Genetic Analyzer to obtain a DNA profile and sex confirmation. The successful extraction of mtDNA from teeth using kits already available on the market could help with cost efficiency in laboratories. Future goals of this study include applying extraction methods on teeth and other remains that range in age from years to decades to centuries old. Ultimately the goal is to design a method for extraction and analysis of DNA from ancient remains.
References: Dawson Cruz T, Thomas JT, Berlin RM, Barker JM. (2013). Advanced Forensic DNA Analysis: Mitochondrial DNA Amplification. Department of Forensic Science, Virginia Commonwealth University. doi: 10.1111/1556-4029.12171.
Powell Jr, Thomas E. ‘Mitochondrial DNA Polymorphisms in Human Evolution DNA Extraction and Amplification Kit with 0.2-ML Tubes.’ Carolina Biological Supply: World-Class Support for Science & Math, Science and Technology Concepts’ Program and the Smithsonian Science Education Center.
Funder Acknowledgement(s): I would like to thank Ms. Jacquiline Vinson, program coordinator of the Louis Stokes Mississippi Alliance for Minority Participation. In addition, I would like to thank Dr. Murrell Godfrey and Caroline Spencer for their guidance throughout this science investigation thus far.
Faculty Advisor: Jacquiline Vinson, email@example.com
Role: I played a role in all parts of this research project.