Getting Ahead: Extraction of DNA from Skeletonized Cranial Material and Teeth

Area of Science:

• Forensic Anthropology
• Molecular Biology
• Genetics

Background:

• The Defense POW/MIA Accounting Agency (DAPMA) handles identification of missing U.S. service members.
• Over 2177 cranial elements and 1565 teeth were submitted for DNA testing between 1990 and 2018.
• Recovered materials span World War II, Korean War, and Southeast Asia conflicts.

Purpose of the Study:

• To establish optimal DNA testing strategies for skeletonized cranial materials.
• To improve the identification success rate of missing United States service members.
• To compare the efficacy of different DNA extraction and sequencing methods on various skeletal elements.

Main Methods:

• Utilized four distinct DNA extraction protocols.
• Employed mitochondrial DNA Sanger sequencing, modified AmpFlSTR® Yfiler™, AmpFlSTR® MiniFiler™, PowerPlex® Fusion, and Next Generation Sequencing.
• Analyzed DNA from cranial elements and teeth recovered from various conflict eras.

Main Results:

• Cranial elements showed most consistent results with Sanger sequencing using organic purification.
• Teeth were most successful for Sanger sequencing with inorganic purification.
• STR testing yielded inverse results for cranial bones compared to Sanger sequencing.
• Temporal cranial elements provided the most consistent results among all cranial samples.

Conclusions:

• Specific DNA extraction and purification methods are crucial for different skeletal elements.
• Sanger sequencing is optimal for cranial elements with organic purification, while teeth benefit from inorganic purification.
• STR testing strategies should be adapted based on the skeletal source material for improved identification outcomes.