%0 Journal Article %A Diaz-Cano, Salvador J. %A P Brady, S %D 2012 %T DNA extraction from formalin-fixed, paraffin-embedded tissues: protein digestion as a limiting step for retrieval of high-quality DNA %U https://figshare.com/articles/journal_contribution/DNA_extraction_from_formalin-fixed_paraffin-embedded_tissues_protein_digestion_as_a_limiting_step_for_retrieval_of_high-quality_DNA/97660 %R 10.6084/m9.figshare.97660.v1 %2 https://ndownloader.figshare.com/files/223965 %K DNA extraction %K molecular technique %K paraffin %K paraffinembedded %K protein digestion %K Molecular Biology %K Medicine %K Cancer %X

Several DNA extraction methods have been used for formalin-fixed, 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 micrograms/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 beta-globin gene tested the suitability of the retrieved DNA for PCR amplification.

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