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Submitted
March 10, 2023
Published
December 23, 2022
Effect of extreme heat on dna obtained from tooth of burnt corpses
Corresponding Author(s) : Leena Kumari
leena.bhardwaj2000@gmail.com
International Journal of Allied Medical Sciences and Clinical Research,
Vol. 10 No. 4 (2022): 2022 Volume -10 - Issue 4
Abstract
Introduction: Identification of sex has a crucial role in personal identification. This study aims to determine the sex of an individual from burnt teeth samples exposed to a burning temperature of 2500C for 15minutes and 45 minutes respectively through RT-PCR on amelogenin sex marker.
A total of 60 tooth samples were subjected to the burning temperature at 2500C for 15minutes and 45 minutes respectively. The tooth was grinded with tissue lyzer and DNA extraction was done by the phenol-chloroform method. All tooth samples were quantified for DNA concentration and then analyzed with RT-PCR
Although there are various methods of DNA isolation and extraction from teeth, yet, comparison across these methods to measure the effectiveness of the specific method is not clearly established. Thus, the primary objective of this study was to measure the effectiveness across the grinding and broaching methods of DNA extraction in freshly extracted tooth pulp.Real Time-Polymerase Chain Reaction (RT-PCR) and subsequent multiplex STR typing.
Method: A total of 40 freshly extracted normal teethwere randomly collected.Isolation and extraction of DNA was doneby organic extraction method. Precipitation of samples was done using 100% chilled ethyl alcohol followed by concentration and washing of DNA via column based techniqueusing DNA binding buffer and DNA wash buffer. Agarose gel electrophoresis was done to roughly estimate the DNA content while exact quantity of DNA was estimated by RT-PCR technique.
Result: Independent sample t test analysis revealed that the mean quantity of DNA (in ?g/l) was significantly higher in broaching method (M=29.91, SD=0.65) than grinding (M=9.71, SD=0.45), t(38)=114.19, p<0.000. Similarly, quality of DNA was analyzed using smear quality and it was found that the quality of DNA for broaching (M=4.55, SD=0.51) was significantly higher that the grinding method (M=2.55, SD=0.6), t(38)=11.3, p<0.000.
Conclusion: The significant quantitative and qualitative loss of DNAwas observed intooth samples exposed to processed via grinding method compared to broaching method which was further supported by the fact that DNA profiles generated from the DNA extracted using broaching method provided adequate resolution of the autosomal markers andsex identification marker (amelogenin marker) in the present study which isvaluable for human identification and the gender identification finallyleading to individualization.
A total of 60 tooth samples were subjected to the burning temperature at 2500C for 15minutes and 45 minutes respectively. The tooth was grinded with tissue lyzer and DNA extraction was done by the phenol-chloroform method. All tooth samples were quantified for DNA concentration and then analyzed with RT-PCR
Although there are various methods of DNA isolation and extraction from teeth, yet, comparison across these methods to measure the effectiveness of the specific method is not clearly established. Thus, the primary objective of this study was to measure the effectiveness across the grinding and broaching methods of DNA extraction in freshly extracted tooth pulp.Real Time-Polymerase Chain Reaction (RT-PCR) and subsequent multiplex STR typing.
Method: A total of 40 freshly extracted normal teethwere randomly collected.Isolation and extraction of DNA was doneby organic extraction method. Precipitation of samples was done using 100% chilled ethyl alcohol followed by concentration and washing of DNA via column based techniqueusing DNA binding buffer and DNA wash buffer. Agarose gel electrophoresis was done to roughly estimate the DNA content while exact quantity of DNA was estimated by RT-PCR technique.
Result: Independent sample t test analysis revealed that the mean quantity of DNA (in ?g/l) was significantly higher in broaching method (M=29.91, SD=0.65) than grinding (M=9.71, SD=0.45), t(38)=114.19, p<0.000. Similarly, quality of DNA was analyzed using smear quality and it was found that the quality of DNA for broaching (M=4.55, SD=0.51) was significantly higher that the grinding method (M=2.55, SD=0.6), t(38)=11.3, p<0.000.
Conclusion: The significant quantitative and qualitative loss of DNAwas observed intooth samples exposed to processed via grinding method compared to broaching method which was further supported by the fact that DNA profiles generated from the DNA extracted using broaching method provided adequate resolution of the autosomal markers andsex identification marker (amelogenin marker) in the present study which isvaluable for human identification and the gender identification finallyleading to individualization.
Keywords
Amelogenin, DNA, Hydroxyapetite, Gender Identification, Electrophoresis,Forensic Investigation
Leena Kumari, Tirath Das Dogra, Rakesh Dube, Vijay Kumar, Braja Kishore Mohapatra, & Kamal Chauhan. (2022). Effect of extreme heat on dna obtained from tooth of burnt corpses. International Journal of Allied Medical Sciences and Clinical Research, 10(4), 506–513. https://doi.org/10.61096/ijamscr.v10.iss4.2022.506-513
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References
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1. Gunn A. Essential forensic biology. John Wiley & Sons; 2019.
2. Higgins D, Austin JJ. Teeth as a source of DNA for forensic identification of human remains: a review. Sci Justice. 2013;53(4):433-41. doi: 10.1016/j.scijus.2013.06.001, PMID 24188345.
3. Manjunath BC, Chandrashekar BR, Mahesh M, Vatchala Rani RM. DNA profiling and forensic dentistry–A review of the recent concepts and trends. J Forensic Leg Med. 2011;18(5):191-7. doi: 10.1016/j.jflm.2011.02.005, PMID 21663865.
4. Datta P, Datta SS. Role of deoxyribonucleic acid technology in forensic dentistry. J Forensic Dent Sci. 2012;4(1):42-6. doi: 10.4103/0975-1475.99165, PMID 23087582.
5. Hervella M, Iñiguez MG, Izagirre N, Anta A, de-la-Rúa C. Nondestructive methods for recovery of biological material from human teeth for DNA extraction. J Forensic Sci. 2015;60(1):136-41. doi: 10.1111/1556-4029.12568, PMID 25047360.
6. Nelson SJ. Wheeler’s dental anatomy, physiology and occlusion-e-book. Elsevier Health Sciences; 2014.
7. Jeffreys A, Wilson V, Thein S. Hypervariable'minisatellite'regions in human DNA. Biotechnology (Reading, MA). 1985; 467: 1992;24.
8. Butler JM. Short tandem repeat typing technologies used in human identity testing. BioTechniques. 2007;43(4):Sii-Sv:ii-v. doi: 10.2144/000112582, PMID 18019344.
9. Alia-García E, Parra-Pecharromán D, Sánchez-Díaz A, Mendez S, Royuela A, Gil-Alberdi L, et al. Forensic identification in teeth with caries. Forensic Sci Int. 2015;257:236-41. doi: 10.1016/j.forsciint.2015.08.021, PMID 26386340.
10. Devaraju, Rama Raju; Gantala, Ramlal; Ambati et al. DNA detection in tooth exposed to different temperatures: an in vitro study. Journal of Indian Academy of Oral Medicine and Radiology 26(4):p 393-397, Oct–Dec 2014. doi: 10.4103/0972-1363.155681
11. Zagga AD Zagga AD. Forensic study for genetic sex determination of burnt powdered skeletal fragments from Sokoto, Northwestern Nigeria. Journal of Dental and Medical Sciences. 2013; 7(5):47-54 doi:10.9790/0853-0754754
References
1. Gunn A. Essential forensic biology. John Wiley & Sons; 2019.
2. Higgins D, Austin JJ. Teeth as a source of DNA for forensic identification of human remains: a review. Sci Justice. 2013;53(4):433-41. doi: 10.1016/j.scijus.2013.06.001, PMID 24188345.
3. Manjunath BC, Chandrashekar BR, Mahesh M, Vatchala Rani RM. DNA profiling and forensic dentistry–A review of the recent concepts and trends. J Forensic Leg Med. 2011;18(5):191-7. doi: 10.1016/j.jflm.2011.02.005, PMID 21663865.
4. Datta P, Datta SS. Role of deoxyribonucleic acid technology in forensic dentistry. J Forensic Dent Sci. 2012;4(1):42-6. doi: 10.4103/0975-1475.99165, PMID 23087582.
5. Hervella M, Iñiguez MG, Izagirre N, Anta A, de-la-Rúa C. Nondestructive methods for recovery of biological material from human teeth for DNA extraction. J Forensic Sci. 2015;60(1):136-41. doi: 10.1111/1556-4029.12568, PMID 25047360.
6. Nelson SJ. Wheeler’s dental anatomy, physiology and occlusion-e-book. Elsevier Health Sciences; 2014.
7. Jeffreys A, Wilson V, Thein S. Hypervariable'minisatellite'regions in human DNA. Biotechnology (Reading, MA). 1985; 467: 1992;24.
8. Butler JM. Short tandem repeat typing technologies used in human identity testing. BioTechniques. 2007;43(4):Sii-Sv:ii-v. doi: 10.2144/000112582, PMID 18019344.
9. Alia-García E, Parra-Pecharromán D, Sánchez-Díaz A, Mendez S, Royuela A, Gil-Alberdi L, et al. Forensic identification in teeth with caries. Forensic Sci Int. 2015;257:236-41. doi: 10.1016/j.forsciint.2015.08.021, PMID 26386340.
10. Devaraju, Rama Raju; Gantala, Ramlal; Ambati et al. DNA detection in tooth exposed to different temperatures: an in vitro study. Journal of Indian Academy of Oral Medicine and Radiology 26(4):p 393-397, Oct–Dec 2014. doi: 10.4103/0972-1363.155681
11. Zagga AD Zagga AD. Forensic study for genetic sex determination of burnt powdered skeletal fragments from Sokoto, Northwestern Nigeria. Journal of Dental and Medical Sciences. 2013; 7(5):47-54 doi:10.9790/0853-0754754
2. Higgins D, Austin JJ. Teeth as a source of DNA for forensic identification of human remains: a review. Sci Justice. 2013;53(4):433-41. doi: 10.1016/j.scijus.2013.06.001, PMID 24188345.
3. Manjunath BC, Chandrashekar BR, Mahesh M, Vatchala Rani RM. DNA profiling and forensic dentistry–A review of the recent concepts and trends. J Forensic Leg Med. 2011;18(5):191-7. doi: 10.1016/j.jflm.2011.02.005, PMID 21663865.
4. Datta P, Datta SS. Role of deoxyribonucleic acid technology in forensic dentistry. J Forensic Dent Sci. 2012;4(1):42-6. doi: 10.4103/0975-1475.99165, PMID 23087582.
5. Hervella M, Iñiguez MG, Izagirre N, Anta A, de-la-Rúa C. Nondestructive methods for recovery of biological material from human teeth for DNA extraction. J Forensic Sci. 2015;60(1):136-41. doi: 10.1111/1556-4029.12568, PMID 25047360.
6. Nelson SJ. Wheeler’s dental anatomy, physiology and occlusion-e-book. Elsevier Health Sciences; 2014.
7. Jeffreys A, Wilson V, Thein S. Hypervariable'minisatellite'regions in human DNA. Biotechnology (Reading, MA). 1985; 467: 1992;24.
8. Butler JM. Short tandem repeat typing technologies used in human identity testing. BioTechniques. 2007;43(4):Sii-Sv:ii-v. doi: 10.2144/000112582, PMID 18019344.
9. Alia-García E, Parra-Pecharromán D, Sánchez-Díaz A, Mendez S, Royuela A, Gil-Alberdi L, et al. Forensic identification in teeth with caries. Forensic Sci Int. 2015;257:236-41. doi: 10.1016/j.forsciint.2015.08.021, PMID 26386340.
10. Devaraju, Rama Raju; Gantala, Ramlal; Ambati et al. DNA detection in tooth exposed to different temperatures: an in vitro study. Journal of Indian Academy of Oral Medicine and Radiology 26(4):p 393-397, Oct–Dec 2014. doi: 10.4103/0972-1363.155681
11. Zagga AD Zagga AD. Forensic study for genetic sex determination of burnt powdered skeletal fragments from Sokoto, Northwestern Nigeria. Journal of Dental and Medical Sciences. 2013; 7(5):47-54 doi:10.9790/0853-0754754
