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Anthony Killeen has assembled a collection of readily reproducible molecular pathology techniques that are either frequently performed or recognized for their significant diagnostic utility. Each method is described in step-by-step detail by a leading molecular pathologist or laboratory scientist who has developed it or used it extensively. These clinical laboratory techniques can be used for the diagnosis or monitoring of cancer, hematological malignancies, infectious diseases, and selected genetic disorders. Comprehensive and path-breaking, Molecular Pathology Protocols will enable clinical laboratories to introduce new molecular pathology tests and lay the groundwork for a much-needed standardization in this rapidly developing field.
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M E T H O D S I N M O L E C U L A R M E D I C I N E TM Molecular Pathology Protocols Edited by Anthony A. Killeen Humana Press DNA Extraction from Paraffin-Embedded Tissues 1 1 DNA Extraction from Paraffin-Embedded Tissues Hongxin Fan and Margaret L. Gulley 1. Introduction In routine histopathology, most tissues are fixed in formalin and embedded in paraffin for long-term preservation. DNA can be extracted from these tissues for subsequent molecular analysis by amplification methods. We describe herein a protocol for DNA preparation from paraffin-embedded tissues based on published procedures (1–3). In brief, tissue sections are placed into microfuge tubes, then deparaffinized with xylene. The xylene is removed with ethanol washes, and the tissue is treated with proteinase K to make DNA available for amplification. This protocol is simple, but there are several factors that influence the success of subsequent DNA amplification assays, including the type of fixative that is used, the duration of fixation, the age of the paraffin block, and the length of the DNA segment to be