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Extraction of DNA from Skeletonized Postcranial Remains: A Discussion of Protocols and Testing Modalities
Suni M Edson1,2
1Armed Forces DNA Identification Laboratory, Armed Forces Medical Examiner System, 115 Purple Heart Drive, Dover AFB, DE, 19902.
Journal of Forensic Sciences
|March 30, 2019
Summary
This study reviewed DNA analysis of over 13,000 remains from WWII, Korean War, and Southeast Asia. Complete demineralization with organic purification optimized mitochondrial DNA sequencing, while inorganic purification was best for STR testing.
Area of Science:
- Forensic Science
- Molecular Biology
- Genetics
Background:
- The Armed Forces DNA Identification Laboratory (AFDL) has conducted extensive DNA analysis for identifying remains.
- Over 13,000 cases involving wartime losses from World War II, Korean War, and Southeast Asia were analyzed between 1990 and 2018.
Purpose of the Study:
- To retrospectively analyze DNA analysis methods used by the AFDL.
- To evaluate the effectiveness of different DNA extraction protocols for various DNA analyses on postcranial osseous materials.
Main Methods:
- Analysis of over 13,000 postcranial osseous samples using mitochondrial DNA sequencing, AmpFlSTR® Yfiler™, AmpFlSTR® MiniFiler™, PowerPlex® Fusion, and Next-Generation Sequencing (NGS).
- Comparison of four DNA extraction protocols: incomplete demineralization with organic purification, complete demineralization with organic purification, complete demineralization with inorganic purification (QIAquick PCR Purification Kit), and an NGS-specific protocol.
Main Results:
- Complete demineralization coupled with organic purification was generally optimal for mitochondrial DNA sequencing across all tested osseous elements.
- Demineralization paired with inorganic purification yielded optimal results for Short Tandem Repeat (STR) testing, irrespective of the kit or osseous element.
Conclusions:
- The choice of DNA extraction protocol significantly impacts the success of downstream DNA analysis.
- Specific protocols are recommended for optimal results in mitochondrial DNA sequencing versus STR analysis in forensic casework involving ancient or degraded osseous remains.
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