DNA Extraction from Formalin-fixed, Paraffin-embedded Tissues: Protein Digestion as a Limiting Step for Retrieval of High-quality DNA


    loading  Checking for direct PDF access through Ovid

Abstract

Several DNA extraction methods have been used for formalinfixed, paraffin-embedded tissues, with variable results being reported regarding the suitability of DNA obtained from such sources to serve as template in polymerase chain reaction (PCR)-based genetic analyses. We present a method routinely used for archival material in our laboratory that reliably yields DNA of sufficient quality for PCR studies. This method is based on extended proteinase K digestion (250 μg/ml in an EDTA-free calcium-containing buffer supplemented with mussel glycogen) followed by phenol-chloroform extraction. Agarose gel electrophoresis of both digestion buffer aliquots and PCR amplification of the β-globin gene tested the suitability of the retrieved DNA for PCR amplification.

    loading  Loading Related Articles