A Powder-free DNA Extraction Workflow for Skeletal Samples.

Area of Science:

• Forensic science
• Molecular biology
• Anthropology

Background:

• Skeletal material processing for DNA extraction presents significant challenges, including labor intensity, time consumption, specialized equipment needs, and contamination risks.
• Current protocols often require powdering bone or teeth, which adds complexity and time to the workflow.

Purpose of the Study:

• To evaluate a non-powdering DNA extraction method for skeletal samples using the PrepFiler® BTA™ Forensic DNA Extraction Kit.
• To optimize sample mass and incubation times for efficient DNA recovery from whole bone chips.
• To assess the effectiveness of this method on diverse skeletal remains from human cadavers.

Main Methods:

• Testing various sample masses (e.g., 50 mg, 150 mg) and incubation times (2, 4, 16 hours) with whole bone chips.
• Applying the optimized method to bones and tooth fragments from human cadavers using an automated platform.
• Comparing results with traditional powdering methods for challenging samples.

Main Results:

• Over 33% of skeletal samples yielded full DNA profiles without powdering or protocol modification.
• Incomplete short tandem repeat (STR) profiles were more common in non-powdered samples with low DNA yields.
• Powdered tissue provided slightly improved results for samples with limited DNA.

Conclusions:

• A faster, non-powdering DNA extraction method shows promise for processing skeletal samples.
• This approach can serve as an effective first-pass screening tool in forensic laboratories.
• Further optimization may be needed for samples with very low DNA quantities.