Recovery of High Molecular Weight DNA from Old viscera Samples via Two Established Methods for Human Identification: A Comparative Study
DOI:
https://doi.org/10.48165/Keywords:
Viscera, PCR amplification, DNA profiling, Organic extraction, Extraction using EZ1 bio-robotAbstract
At autopsy, in cases of suspected poisoning or drug intoxication, the viscera are mostly preserved in saturated common salt solution for toxicological analysis. However, frequent situation arises where the preserved viscera sample gets questioned for its source of origin and identification of the individual contributing the said viscera becomes important. In such a scenario, to identify the source of viscera, the DNA Profiling technique can be employed. This study aims at recovery of high molecular weight DNA suitable for DNA profiling from a 5-year-old viscera sample through modified validated methods. Approximately 2 g of viscera samples (kidney) were taken for the study and two different established DNA extraction methods were employed for isolating DNA for downstream procedures viz. quantitation of the extracted DNA, the amplification of the desired loci Polymerase chain reaction (PCR) and its subsequent genotyping. The yield of DNA extracted from both the methods was quantitated using real time PCR system and a complete DNA profile was obtained for the purpose of comparison with the biological relatives of the individual.
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References
[1] Bomjen G, Raina A, Sulaiman IM, Hasnain SE, Dogra TD. Effect of various storage conditions of human tissues on DNA fingerprinting. Journal for Medical Toxicology 1994;14(1):1–6.
[2] Hagelberg E, Gray IC, Jeffreys AJ. Identification of the skeletal remains of a murder victim by DNA analysis. Nature 1991;352:427–429.
[3] Iwamura M, Sadayo E. Analysis of human DNA bone: qualitative study of compact bone microstructure. http:/
/www.openthesis.org/documents/Analysis-human DNA-bone-qualitative-485260.html
[4] Jakubowska J, Maciejewska A, Pawlowski R. Comparison of three methods of DNA extraction from human bones with different degrees of degradation. International Journal of Legal Medicine 2012;126(2):173–178.
[5] Alonso A, Martín P, Albarrán C, García P, García O, Simón LF, et al. Real-time PCR designs to estimate nuclear and mitochondrial DNA copy number in forensic and ancient DNA studies. Forensic Science International 2004;139(1):141–149.
[6] Niederstätter H, Köchl S, Grubwieser P, Pavlic M, Steinlechner M, Parson W. A modular real-time PCR concept for determining the quantity and quality of human nuclear and mitochondrial DNA. Forensic Science International: Genetics 2007;1:29–34.
[7] Dogra TD, Lalwani S, Millo T, Pooniya S, Raina A. Quality and quantity of extracted deoxyribonucleic acid (DNA) from preserved soft tissues of putrefied unidentifiable human corpse. Journal of Laboratory Physicians 2014;6(1):31–35.
[8] Lee SB, McCord B, Buel E. Advances in forensic DNA quantification: a review. Electrophoresis 2014;35(21– 22):3044–3052.
[9] Michaud CL, Foran DR. Simplified field preservation of tissues for subsequent DNA analyses. Journal of Forensic Science 2011;56(4):846–852.
[10] Witt S, Neumann J, Zierdt H, Gébel G, Röscheisen C. Establishing a novel automated magnetic bead-based method for extraction of DNA from a variety of Forensic samples. Forensic Science International 2012;6(2):539– 547.
[11] Butler JM. Forensic DNA typing—biology, technology, and genetics of STR markers. Elsevier Academic Press; 2005. pp. 33–412.
