Forensic Science International: Genetics
Volume 4, Issue 2 , Pages 80-88, February 2010

Optimized manual and automated recovery of amplifiable DNA from tissues preserved in buffered formalin and alcohol-based fixative

  • Kristin Duval

      Affiliations

    • Royal Canadian Mounted Police, National Services and Research, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada
    • ,
  • Rémy A. Aubin

      Affiliations

    • Health Canada, Biologics and Genetic Therapies Directorate, 251 Sir F. G. Banting Drive, Ottawa, Ontario K1A 0K9, Canada
    • Corresponding Author InformationCorresponding author. Tel.: +1 613 954 6468; fax: +1 613 941 8933.
    • ,
  • James Elliott

      Affiliations

    • Royal Canadian Mounted Police, National Services and Research, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada
    • ,
  • Ivan Gorn-Hondermann

      Affiliations

    • Ottawa Hospital Regional Cancer Centre, Ottawa, Ontario K1H 1C4, Canada
    • ,
  • H. Chaim Birnboim

      Affiliations

    • Ottawa Hospital Regional Cancer Centre, Ottawa, Ontario K1H 1C4, Canada
    • ,
  • Derek Jonker

      Affiliations

    • Ottawa Hospital Regional Cancer Centre, Ottawa, Ontario K1H 1C4, Canada
    • ,
  • Ron M. Fourney

      Affiliations

    • Royal Canadian Mounted Police, National Services and Research, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada
    • ,
  • Chantal J. Frégeau

      Affiliations

    • Royal Canadian Mounted Police, National Services and Research, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada
    • Corresponding Author InformationCorresponding author. Tel.: +1 613 993 4206; fax: +1 613 952 7325.

Received 10 March 2009; received in revised form 12 May 2009; accepted 6 June 2009. published online 09 July 2009.

Abstract 

Archival tissue preserved in fixative constitutes an invaluable resource for histological examination, molecular diagnostic procedures and for DNA typing analysis in forensic investigations. However, available material is often limited in size and quantity. Moreover, recovery of DNA is often severely compromised by the presence of covalent DNA–protein cross-links generated by formalin, the most prevalent fixative. We describe the evaluation of buffer formulations, sample lysis regimens and DNA recovery strategies and define optimized manual and automated procedures for the extraction of high quality DNA suitable for molecular diagnostics and genotyping. Using a 3-step enzymatic digestion protocol carried out in the absence of dithiothreitol, we demonstrate that DNA can be efficiently released from cells or tissues preserved in buffered formalin or the alcohol-based fixative GenoFix™. This preparatory procedure can then be integrated to traditional phenol/chloroform extraction, a modified manual DNA IQ™ or automated DNA IQ™/Te-Shake™-based extraction in order to recover DNA for downstream applications. Quantitative recovery of high quality DNA was best achieved from specimens archived in GenoFix™ and extracted using magnetic bead capture.

Keywords: DNA extraction, Automation, Formalin, Alcohol fixation, Magnetic beads, Genotyping, STR, PCR

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PII: S1872-4973(09)00093-3

doi:10.1016/j.fsigen.2009.06.003

Forensic Science International: Genetics
Volume 4, Issue 2 , Pages 80-88, February 2010

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