Polymerase Chain Reaction Analysis of DNA from Paraffin-Embedded Tissue
| Abstract |
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One of the greatest potentials of polymerase chain reaction (PCR) lies in the fact that even minute amounts of target DNA
or extensively damaged DNA can be successfully amplified in vitro and thus become amenable to further study. This enables
a detailed molecular analysis of small amounts of DNA from tissue that has been damaged by fixation (e.g., in formalin) and
long-term storage in paraffin. The applications of this methodology are nearly unlimited. For example, rare tumors that are
stored as formalin-fixed, paraffin-embedded tissue in pathology departments throughout the world can be analyzed at the molecular
level. Furthermore, tissue from small lesions (e.g., primary skin melanomas), which are only rarely available for molecular
analysis since the entire specimen is usually needed for histopathological assessment, can be examined. For PCR analysis,
only several sections from the paraffin block, which are usually dispensable, are sufficient. Even small amounts of very low
quality DNA can be used, as the sensitivity of the detection of specific target DNA sequences is several orders of magnitude
higher than that with any conventional method. For example, Southern blot analysis of DNA from paraffin-embedded tissue has
been performed, but with limited success as only relatively small amounts of degraded and irreversibly modified DNA can be
obtained from embedded specimens (1,2). Thus, PCR methodology creates an ideal link between traditional histology and modern molecular biology (3).
Affiliation(s): (2) Department of Dermatology, University of Munich, Germany
(3) Cornell University Graduate School of Medical Sciences and the Laboratory of Molecular Pharmacology, Memorial Sloan-Kettering Cancer Center, New York
(4) Memorial Sloan-Kettering Cancer Center, New York
(3) Cornell University Graduate School of Medical Sciences and the Laboratory of Molecular Pharmacology, Memorial Sloan-Kettering Cancer Center, New York
(4) Memorial Sloan-Kettering Cancer Center, New York
Book Title: PCR Protocols: Current Methods and Applications
Series: Methods in Molecular Biology | Volume: 15 | Pub. Date: Feb-03-1993 | Page Range: 81-88 | DOI: 10.1385/0-89603-244-2:81
Subject: Genetics/Genomics
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