DNA extraction from formalin-fixed, paraffin-embedded tissues: protein digestion as a limiting step for retrieval of high-quality DNA

Diagn Mol Pathol. 1997 Dec;6(6):342-6. doi: 10.1097/00019606-199712000-00006.

Abstract

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.

MeSH terms

  • Adrenal Glands / chemistry
  • Breast / chemistry
  • Colon / chemistry
  • DNA / isolation & purification*
  • Formaldehyde / chemistry*
  • Humans
  • Hydrolysis
  • Paraffin Embedding / methods*
  • Proteins / genetics*
  • Proteins / metabolism
  • Skin / chemistry
  • Thyroid Gland / chemistry
  • Tissue Fixation / methods
  • Urinary Bladder / chemistry

Substances

  • Proteins
  • Formaldehyde
  • DNA