Date of Submission
5-2024
Document Type
Thesis
Degree Name
Master of Science in Forensic Science
Department
Forensic Science
Advisor
David San Pietro, Ph.D.
Committee Member
Ashley Morgan, Ph.D.
Committee Member
Wendy Gruhl, M.S.
Keywords
DNA Degradation, DNA Repair Mechanisms, Heat Damage
MeSH
DNA Repair, DNA Damage
LCSH
DNA repair, DNA damage
Abstract
The effects of DNA degradation via temperature on potential DNA repair mechanisms were assessed. DNA was extracted from known whole blood samples as well as whole blood stains placed on cotton swatches. The swatches were placed into an oven and subjected to varying temperatures (ranges from 50̊ C to 200̊ C) for different periods of time (ranging from 0 to to 12 hours) to induce degradation prior to subsequent extraction with the QIAGEN© QIAamp DNA Investigator Kit, quantification with Quantifiler Trio DNA Quantification Kit, amplification with Globalfiler PCR Amplification Kit, and capillary electrophoresis with Applied Biosystems 3500xl Genetic Analyzer, Applied Biosystems 3500 Series Data Collection Software 4, and GeneMarker® analysis software. Half of the samples from each set of swatches were repaired with New England BioLabs© PreCR ® Repair Mix while the others were left without repair. It was found that peak heights increased in response to repair mechanisms while those loci where alleles fully dropped out did not. The repair mixture appeared to repair only those remaining alleles after degradation. It was also seen that there was a higher potential for contamination in samples degraded and repaired for longer periods of time, due to possible drop-in. This is a reliable method and there is the potential for casework; although, further research should be done to get the full scope of repair situations. Data will be presented pertaining to mixtures samples degraded and repaired under these conditions to preliminarily assess its effect on both peak height and mixture ratios.
Recommended Citation
Leffler, Sydney M., "Effect of Heat-Induced Damage on the Efficiency of Possible DNA Repair Mechanisms" (2024). Master's Theses. 220.
https://digitalcommons.newhaven.edu/masterstheses/220