P195 – Assessing DNA Yield in Three Bones Sites Using a Less Destructive Powdering Method

11:13

Thursday August 20^th

Station 07

Duration: 12 minutes

03. Human identification

Juliana Bablak

In many forensic contexts, human remains are the only DNA source that is left from a crime scene, making genetic analysis of skeletal material especially for human identification. Bone sampling methods commonly involve cutting the bone, potentially compromising bone integrity. In situations where ethical concerns and respect for human remains are crucial, evaluating less destructive alternatives is necessary.

This research investigates the variation of DNA yield from cortical bone, trabecular bone, and the medullary cavity of five femurs using a minimally destructive powdering method. Three replicates from each anatomical site were collected using a Dremel tool for minimal damage to the skeletal remains, obtaining approximately 100 mg of bone powder. For the medullary cavity and the trabecular sites, holes of about 10mm were made, while for the cortical site, powder from scraping was obtained. DNA was extracted using a modified EZ1 protocol optimized for bone samples. DNA quantity, degradation, and genotyping efficiency were assessed using qPCR and STR amplification. To further test sampling efficiency, a subset of samples from two femurs was sequenced for ForenSeq DNA Signature Prep Kit on the Illumina MiSeq platform.

Results indicate that cortical bone has significantly higher DNA concentrations than the medullary cavity and trabecular bone, with an average of 23 times more DNA, and full or partial STR profiles were obtained for cortical samples. On the contrary, the medullary cavity bone samples produced minimal DNA yields and limited genetic profiles. Trabecular bone produced little to no results with concentrations below 0.06ng/ul (in the 91bp autosomal amplicon region). Next-generation sequencing data similarly demonstrated a decrease in both STR and SNPs marker recovery from cortical bone to medullary cavity and trabecular bone.

It is important to highlight that the human remains used in this study were donated and analyzed immediately after defleshing and cleaning, without prolonged exposure to environmental conditions or other stressors that could potentially affect DNA preservation. Future research focusing on older and environmentally exposed remains will be essential to determine whether this minimally destructive sampling strategy is suitable for challenged forensic samples.

Authors

Juliana Bablak (Towson Univeristy, United States of America)
Filipa Simao (Towson University, United States of America)

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P195 – Assessing DNA Yield in Three Bones Sites Using a Less Destructive Powdering Method

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