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)