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DNA Extraction from Blood and Forensic Samples

  • Solange Sorçaburu Cigliero
  • Elisabetta Edalucci
  • Paolo Fattorini
Chapter

Abstract

Genetic analyses are performable from all the biological samples containing DNA. Thus, DNA extraction is the first and probably one of the most crucial steps of any genetic test. This chapter provides protocols for DNA extraction from both fresh blood and the big variety of biological samples that can be studied in the Forensic Laboratory. First, the general precautions which have to be adopted to prevent cross-contamination of the samples are underlined. All the extraction protocols (based upon Proteinase K/SDS lysis followed by ethanol precipitation) are carefully described while crucial steps are pointed out by footnotes. In addition, since PCR failure is common when handling aged/forensic specimens, strategies to remove substances acting as PCR inhibitors are described as well. At least, a simple and inexpensive PCR-based method allowing discriminating between removable and un-removable PCR inhibition is shown.

Keywords

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These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Alaeddini R, Walsh SJ, Abbas A (2010) Forensic implications of genetic analyses from degraded DNA – a review. Forensic Sci Int Genet 4:148–157PubMedCrossRefGoogle Scholar
  2. 2.
    Paabo S, Poinar H, Serre D, Jaenicke-Despres V, Hebler J, Rohland N, Kuch M, Krause J, Vigilant L, Hofreiter M (2004) Genetic analyses from ancient DNA. Annu Rev Genet 38:645–679PubMedCrossRefGoogle Scholar
  3. 3.
    Eckert KA, Kunkel TA (1991) in McPherson MJ, Quirke P, Taylor GR (Eds) PCR: a practical approach IRL Press, Oxford, 225–244Google Scholar
  4. 4.
    Allouche M, Hamdoum M, Mangin P, Castella V (2008) Genetic identification of decomposed cadavers using nails as DNA source. Forensic Sci Int Genet 3(1):46–49PubMedCrossRefGoogle Scholar
  5. 5.
    Rohland N, Hofreiter M (2007) Ancient DNA extraction from bones and teeth. Nat Protoc 2(7):1756–1762PubMedCrossRefGoogle Scholar
  6. 6.
    Rasmussen M, Li Y, Lindgreen S, Pedersen JS, Albrechtsen A, Moltke I, Metspalu M, Metspalu E, Kivisild T, Gupta R, Bertalan M, Nielsen K, Gilbert MT, Wang Y, Raghavan M, Campos PF, Kamp HM, Wilson AS, Gledhill A, Tridico S, Bunce M, Lorenzen ED, Binladen J, Guo X, Zhao J, Zhang X, Zhang H, Li Z, Chen M, Orlando L, Kristiansen K, Bak M, Tommerup N, Bendixen C, Pierre TL, Gronnow B, Meldgaard M, Andreasen C, Fedorova SA, Osipova LP, Higham TF, Ramsey CB, Hansen TV, Nielsen FC, Crawford MH, Brunak S, Sicheritz-Ponten T, Villems R, Nielsen R, Krogh A, Wang J, Willerslev E (2010) Ancient human genome sequence of an extinct palaeo-eskimo. Nature 463(7282):757–762PubMedCrossRefGoogle Scholar
  7. 7.
    Loreille OM, Diegoli TM, Irwin JA, Coble MD, Parsons TJ (2007) High efficiency DNA extraction from bone by total demineralization. Forensic Sci Int Genet 1(2):191–195PubMedCrossRefGoogle Scholar
  8. 8.
    Fattorini P, Marrubini G, Ricci U, Gerin F, Grignani P, Cigliero SS, Xamin A, Edalucci E, La Marca G, Previderè C (2009) Estimating the integrity of aged DNA samples by CE. Electrophoresis 30(22):3986–3995PubMedCrossRefGoogle Scholar
  9. 9.
    King C, Debruyne R, Kuch M, Schwarz C, Poinar H (2009) A quantitative approach to detect and overcome PCR inhibition in ancient DNA extracts. Biotechniques 47(5):941–949PubMedCrossRefGoogle Scholar
  10. 10.
    Kontanis EJ, Reed FA (2006) Evaluation of real-time PCR amplification efficiencies to detect PCR inhibitors. J Forensic Sci 51(4):795–804PubMedCrossRefGoogle Scholar
  11. 11.
    Hedman J, Nordgaard A, Rasmusson B, Ansell R, Radstrom P (2009) Improved forensic DNA analysis through the use of alternative DNA polymerases and statistical modeling of DNA profiles. Biotechniques 47(5):951–958PubMedCrossRefGoogle Scholar
  12. 12.
    Previderè C, Micheletti P, Perossa R, Grignani P, Fattorini P (2002) Molecular characterisation of the nucleic acids ­recovered from aged forensic samples. Int J Legal Med 116(6):334–339PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  • Solange Sorçaburu Cigliero
    • 1
  • Elisabetta Edalucci
    • 2
  • Paolo Fattorini
    • 1
  1. 1.Department of Medicine, Surgery and HealthUniversity of TriesteTriesteItaly
  2. 2.Department of Nuclear MedicineAzienda U-O di TriesteTriesteItaly

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