Abstract
Formalin fixation is commonly used to preserve tissue sections for pathological testing and embalming cadavers for medical dissection or burial. DNA extracted from formalin-fixed tissues may also provide an alternative source of genetic material for medical diagnosis and forensic casework, such as identifying unknown embalmed human remains. Formaldehyde causes DNA damage, chemical modifications, and degradation, thereby reducing the quantity and quality of DNA available for downstream genetic analyses. By comparing the DNA yield, level of DNA degradation, and short tandem repeat (STR) success of various tissue types, this study is the first of its kind to provide some guidance on which samples from embalmed bodies are likely to generate more complete STR profiles. Tissue samples were dissected from three male embalmed cadavers and included bone, cartilage, hair, muscle, internal organs, skin, teeth, and nail clippings. DNA was purified from all samples using the QIAamp® FFPE Tissue Kit (Qiagen), quantified using the QuantiFiler® Trio DNA Quantification kit (Life Technologies), and genotyped using the GlobalFiler® PCR Amplification Kit (Life Technologies). Results of this study showed variation in DNA quantity and STR success between different types of tissues and some variation between cadavers. Overall, bone marrow samples resulted in the highest DNA yields, the least DNA degradation, and greatest STR success. However, several muscle, hair, and nail samples generated higher STR success rates than traditionally harvested bone and tooth samples. A key advantage to preferentially using these tissue samples over bone (and marrow) and teeth is their comparative ease and speed of collection from the cadaver and processing during DNA extraction. Results also indicate that soft tissues affected by lividity (blood pooling) may experience greater exposure to formalin, resulting in more DNA damage and reduced downstream STR success than tissues under compression. Overall, we recommend harvesting from selected muscles (gastrocnemius, rectus femoris, flexor digitorum brevis, masseter, brachioradialis) or fingernails for human identification purposes.
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Acknowledgments
We would like to thank Samantha Tippen for collecting a subset of the cadaveric tissue samples. We would also like to acknowledge Sheri Olsen at Life Technologies for kindly providing the QuantiFiler® Trio DNA Quantification kit used in this study. The authors would also like to thank the staff at STAFS for their assistance, and the individuals and families of those who donated their bodies to STAFS for scientific research.
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All experiments performed in this research were conducted within the current laws of the USA. Code 45 Code of Federal Regulations part 46102(f) exempts the requirement for Institutional review board (IRB) regarding the use of cadaveric samples.
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Wheeler, A., Czado, N., Gangitano, D. et al. Comparison of DNA yield and STR success rates from different tissues in embalmed bodies. Int J Legal Med 131, 61–66 (2017). https://doi.org/10.1007/s00414-016-1405-5
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DOI: https://doi.org/10.1007/s00414-016-1405-5