Optimized Manual and Automated Recovery of Amplifiable DNA From Tissues Preserved in Buffered Formalin and Alcohol-Based Fixative

• NCJ Number: 229903
• Journal: Forensic Science International: Genetics Volume: 4 Issue: 2 Dated: February 2010 Pages: 80-88
• Author(s): Kristin Duval; Remy A. Aubin; James Elliott; Ivan Gorn-Hondermann; H. Chaim Birnhoim; Derek Jonker; Ron M. Fourney; Chantal J. Fregeau
• Date Published: February 2010
• Length: 9 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: Ireland

Annotation

This article describes 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.

Abstract

Archival tissue preserved in fixative constitutes an invaluable resource for histological examinations, 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. Using a three-step enzymatic digestion protocol carried out in the absence of dithiothreitol, it was demonstrated that DNA can be efficiently released from cells or tissues preserved in buffered formalin or the alcohol-based fixative GenoFixTM. This preparatory procedure can then be integrated to traditional phenol/chloroform extraction, a modified manual DNA IQTM or automated DNA IQTM /Te-ShakeTM -based extraction in order to recover DNA for downstream applications. Quantitative recovery of high-quality DNA was best achieved from specimens archived in GenoFixTM and extracted using magnetic bead capture. Tables, figures, and references (Published Abstract)