13. Extraction and Enzymatic Amplification of DNA from Paraffin-Embedded Specimens
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Histopathology archive material constitutes an enormous resource of diseased tissues. It is composed of specimens that have
usually been fixed with formalin to stop further tissue changes after removal from the body and subsequently embedded in a
supporting material, such as paraffin, allowing sections to be cut for examination by microscopy. It has now been shown that
DNA extracted from formalin-fixed, paraffin-embedded specimens, although degraded to some extent, may be used for molecular
analysis (1–3). Factors determining the size range of the DNA include the length of time between removal of tissue from the patient and
immersion in fixative, the nuclease content of the tissue, and the pH of the formalin. The lower the pH the more fragmented
the DNA, probably owing to depurination. Unfortunately, many histopathology departments routinely use a 10% (v/v) solution
of formaldehyde in water without any buffering agent. Hence, over a period of time, the formaldehyde oxidizes to formic acid
and the pH drops, so that the older the formaldehyde solution the lower the pH. The length of time the specimen has spent
in formalin before processing will also influence the state of the DNA. Fortunately, most small biopsy specimens will have
spent an overnight period or even less in fixative.
Affiliation(s): (2) Institute of Cancer Research, Haddow Laboratories, Sutton, Surrey, UK
(3) Institute of Psychiatry, De Crespigny Park, London, UK
(3) Institute of Psychiatry, De Crespigny Park, London, UK
Book Title: Protocols in Human Molecular Genetics
Series: Methods in Molecular Biology | Volume: 9 | Pub. Date: Nov-01-1991 | Page Range: 133-140 | DOI: 10.1385/0-89603-205-1:133
Subject: Genetics/Genomics
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