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

Forensic Science International. Genetics
Kristin DuvalChantal J Frégeau

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 cap...Continue Reading

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Citations

Jan 24, 2012·Forensic Science International. Genetics·Chantal J Frégeau, Anick De Moors
Nov 27, 2015·BMC Infectious Diseases·Felipe A CastroNicolas Wentzensen
Apr 23, 2013·Analytical Biochemistry·Stefan PairederMargit Cichna-Markl
Jan 27, 2015·Applied Immunohistochemistry & Molecular Morphology : AIMM·Bendik LundHelge Klungland
Dec 15, 2017·BMC Research Notes·Lindsay Gielda, Stefanie Rigg
Apr 11, 2013·Expert Review of Molecular Diagnostics·Serena Bonin, Giorgio Stanta
Nov 8, 2020·Forensic Science, Medicine, and Pathology·Katarina VitoševićDanijela Todorović

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