Abstract
Forensic casework analyses often face challenges, such as limited genetic material with or without fragmentation and damage. To compensate for low amounts and degradation, shorter amplicons are often applied in the analysis. Also, a change of markers might be necessary using mitochondrial instead of autosomal markers. In addition, forensic research often involves analysis of large number of samples for marker evaluation and population-database compilation. Therefore, a flexible, robust but also rapid method for the detection of variation is highly useful. Pyrosequencing® is a rapid, reliable, easy-to-use method for sequence analysis. The method is well suited for rapid forensic analysis of a few targets or analysis of a single target in many samples. It allows sequencing of very short amplicons, which facilitates analysis of degraded DNA. Here we present the use of Pyrosequencing, a robust method for sensitive forensic analysis of mitochondrial DNA, autosomal STRs, and Y-chromosome STRs and SNPs.
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References
Gibbon V, Paximadis M, Strkalj G et al (2009) Novel methods of molecular sex identification from skeletal tissue using the amelogenin gene. Forensic Sci Int Genet 3:74–79
Budowle B, Wilson MR, DiZinno JA et al (1999) Mitochondrial DNA regions HVI and HVII population data. Forensic Sci Int 103:23–35
Lembring M, van Oven M, Montelius M et al (2013) Mitochondrial DNA analysis of Swedish population samples. Int J Legal Med 127:1097–1099
Calloway CD, Reynolds RL, Herrin GL Jr et al (2000) The frequency of heteroplasmy in the HVII region of mtDNA differs across tissue types and increases with age. Am J Hum Genet 66:1384–1397
Cowell RG, Lauritzen SL, Mortera J (2007) Identification and separation of DNA mixtures using peak area information. Forensic Sci Int 166:28–34
Andréasson H, Nilsson M, Budowle B et al (2006) Quantification of mtDNA mixtures in forensic evidence material using pyrosequencing. Int J Legal Med 120:383–390
Andréasson H, Asp A, Alderborn A et al (2002) Mitochondrial sequence analysis for forensic identification using pyrosequencing technology. Biotechniques 32:124–133
Andréasson H, Nilsson M, Styrman H et al (2007) Forensic mitochondrial coding region analysis for increased discrimination using pyrosequencing technology. Forensic Sci Int Genet 1:35–43
Edlund H, Allen M (2009) Y chromosomal STR analysis using Pyrosequencing technology. Forensic Sci Int Genet 3:119–124
Divne AM, Edlund H, Allen M (2010) Forensic analysis of autosomal STR markers using Pyrosequencing. Forensic Sci Int Genet 4:122–129
Kjellström A, Edlund H, Lembring M et al (2012) An analysis of the alleged skeletal remains of Carin Goring. PLoS One 7:e44366
Untergasser A, Cutcutache I, Koressaar T et al (2012) Primer3 new capabilities and interfaces. Nucleic Acids Res 40:e115
Buś MM, Allen M (2014) Collecting and preserving biological samples from challenging environments for DNA analysis. Biopreserv Biobank 12:17–22
Acknowledgements
M.A. received financial support from the Kjell and Märta Beijer Foundation, Vinnova (the Swedish Governmental Agency for Innovation Systems), and Brottsoffermyndigheten (the Swedish Crime Victim Compensation and Support Authority).
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Bus´, M.M., Edlund, H., Allen, M. (2015). Forensic Analysis of Mitochondrial and Autosomal Markers Using Pyrosequencing® . In: Lehmann, U., Tost, J. (eds) Pyrosequencing. Methods in Molecular Biology, vol 1315. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2715-9_26
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DOI: https://doi.org/10.1007/978-1-4939-2715-9_26
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2714-2
Online ISBN: 978-1-4939-2715-9
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