Summary
The highly decomposed remains of a corpse were identified by the amplification and direct sequencing of mitochondrial (mt) DNA. Degraded DNA was extracted from bone fragments and a necrotic skin sample and amplified at 2 hypervariable segments within the mitochondrial non-coding region using 2 rounds of nested PCR. Both strands of the amplified regions were sequenced and compared with each other to ensure fidelity of the data. Sequences from the bone and skin were found to be identical and matched data generated from a blood sample provided by a presumptive sister of the deceased. Thus, this evidence was consistent with the sister and the deceased being related.
Zusammenfassung
Die hochgradig zersetzten Überbleibsel einer Leiche wurden mit Hilfe der Amplifikation und der direkten Sequenzierung der mitochondrialen DNA identifiziert. Degradierte DNA wurde von Knochenfragmenten und von einer nekrotischen Hautprobe extrahiert und in zwei hypervariablen Segmenten der mitochondrialen nicht-kodierenden Region amplifiziert unter Anwendung zweier Runden von „nested” PCR. Beiden Stränge der amplifizierten Regionen wurden sequenziert und miteinander verglichen, um die Vertrauenswürdigkeit der Daten zu sichern. Die Sequenzen von Knochen und von der Haut waren identisch und paßten zu den Daten aus einer Blutprobe, welche von einer mutmaßlichen Schwester des Verstorbenen herrührte. Dieser Beweis war somit konsistent mit der Annahme, daß Schwester und Verstorbene verwandt waren.
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References
Bogenhagen D, Clayton DA (1974) The number of mitochondrial deoxyribonucleic acid genomes in mouse L and human HeLa cells. J Biol Chem 249:7991–7995
Giles RE, Blanc H, Cann HM, Wallace DC (1980) Maternal inheritance of human mitochondrial DNA. Proc Natl Acad Sci USA 77:6715–6719
Anderson S, Bankier AT, Barrell BG, deBruijin MHL, Coulson AR, Drouin J, Eperon IC, Nierlich DP, Roe BA, Sanger F, Schreier PH, Smith AJH, Staden R, Young IG (1981) Sequence and organisation of the human mitochondrial genome. Nature 290:457–465
Aquadro CF, Greenberg BD (1983) Human mitochondrial DNA variation and evolution: analysis of nucleotide sequences from seven individuals. Genetics 103:287–312
Greenberg BD, Newbold JE, Sugino A (1983) Intraspecific nucleotide sequence variability surrounding the origin of replication in human mitochondrial DNA. Gene 21:33–49
Cann RL, Stoneking M, Wilson AC (1987) Mitochondrial DNA and human evolution. Nature 325:457–465
Horai S, Hayasaki K (1990) Intraspecific nucleotide sequence differences in the major noncoding region of human mitochondrial DNA. Am J Hum Genet 46:828–842
Higuchi R, von Beroldingen CH, Sensabaugh GF, Erlich HA (1988) DNA typing from single hairs. Nature 332:543–546
Reynolds R, Sensabaugh G, Blake E (1991) Analysis of genetic markers in forensic DNA samples using the polymerase chain reaction. Anal Chem 63:2–15
Orrego C, King MC (1990) Determination of familial relationships. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR Protocols, Academic Press, London, 416–426
Vigilant L, Pennington R, Harpending H, Kocher TD (1989) Mitochondrial DNA sequences in single hairs from a southern African population. Proc Natl Acad Sci USA 86:9350–9354
Stoneking M, Hedgecock D, Higuchi RG, Vigilant L, Erlich HA (1991) Population variation of human mtDNA control region sequences detected by enzymatic amplification and sequencespecific oligonucleotide probes. Am J Hum Genet 48:370–382
Sullivan KM, Hopgood R, Lang B, Gill P (1991) Automated amplification and sequencing of human mitochondrial DNA. Electrophoresis 12:17–21
Hopgood R, Sullivan KM, Gill P (1992) Strategies for automated sequencing of human mitochondrial DNA directly from PCR products. Biotechniques (in press)
Hagelberg E, Gray IC, Jeffreys AJ (1991) Identification of the skeletal remains of a murder victim. Nature 352:427–429
Hochmeister MN, Budowle B, Borer UV, Eggman U, Comey CT, Dirnhofer R (1991) Typing of deoxyribonucleic acid (DNA) extracted from compact bone from human remains. J Forensic Sci 36:1649–1661
Gill P, Lygo JE, Fowler SJ, Werrett DJ (1987) An evaluation of DNA fingerprinting for forensic purposes. Electrophoresis 8:35–38
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor New York
Di Renzo A, Wilson AC (1991) Branching pattern in the evolutionary tree for human mitochondrial DNA. Proc Natl Acad Sci USA 88:1597–1601
Monnat RJ, Loeb LA (1985) Nucleotide sequence preservation of human mitochondrial DNA. Proc Natl Acad Sci USA 82:2895–2899
Monnat RJ, Reay DT (1986) Nucleotide sequence identity of mitochondrial DNA from different human tissues. Gene 43:205–211
Wallace DC (1991) Mitochondrial DNA mutations and neuromuscular disease. Trends Genet 5:9–13
Gyllensten U, Wharton D, Josefsson A, Wilson AC (1991) Paternal inheritance of mitochondrial DNA in mice. Nature 352:255–257
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Sullivan, K.M., Hopgood, R. & Gill, P. Identification of human remains by amplification and automated sequencing of mitochondrial DNA. Int J Leg Med 105, 83–86 (1992). https://doi.org/10.1007/BF02340829
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DOI: https://doi.org/10.1007/BF02340829