Publication: Comparison of Commercial DNA Extraction Kits for Extraction of DNA From Bones Prepared Using the Novel Theralin Method
Date
2023-11-06
Authors
Bowers, Madalyn
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Abstract
The extraction of DNA from bones to produce high quality samples is extremely important in cases involving unidentified human remains and victims of mass disasters, and many times in these cases, only human bone is available as a source of DNA that can be used to identify the individual. A novel method of preparing bones for extraction using TheralinTM, a precipitating fixative that demineralizes bone, has been developed by researchers at George Mason University. This novel method essentially transforms the bone into a gum-like state that allows for the bone material to be cut into smaller pieces and then digested using lysis buffers. Research has not yet been performed to determine an ideal method of DNA extraction from Theralin-prepared bone, and this study seeks to answer that question. In this study, four different methods of DNA extraction using commercial extraction kits were carried out on bone samples that were prepared using the Theralin method. These extraction methods were the Qiagen QIAamp® DNA Investigator kit protocol with MinElute columns, the PrepFilerTM BTA Forensic DNA Extraction kit protocol with PrepFiler magnetic beads, a combination method using the Qiagen lysis buffers with the PrepFiler magnetic beads, and a combination method using the PrepFiler BTA lysis buffers with the Qiagen MinElute columns. Following extraction with these four methods, the DNA extracts were quantified using the QuantifilerTM Trio DNA Quant kit, and the quantification values of the extracts were compared to determine which method provided the highest amount of DNA. The extracts were amplified using the GlobalFilerTM PCR Amplification kit and genotyped using the Applied Biosystems 3500 Genetic Analyzer with GeneMapperTM ID-X v1.6 software. The STR profiles were compared to the known profiles of the bone samples and analyzed for allelic dropout and signal strength metrics to determine the quality of each DNA extract. The results of this study provided insight into the usefulness and efficiency of the Theralin method of bone preparation, combined with a commercially available DNA extraction technique, to give quality DNA extracts that can be used for downstream applications.
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DNA Extraction, Bones, Theralin