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

Author(s): Díaz-Cano SJ, Brady SP

Publication: Diagn Mol Pathol, 1997, Vol. 6, Page 342-6

PubMed ID: 9559294 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of duration of fixation and proteinase k digestion on PCR success from FFPE specimens. Thyroid, adrenal gland, breast, bladder, and colon specimens fixed for 3-24 h and skin punch biopsies fixed for up to 72 h were compared to fresh specimens.

   Summary of Findings:

   DNA from FFPE specimens was highly degraded compared to DNA from fresh specimens. After proteinase k digestion for 5 days or more, amplification of B-globin was successful, while digestion for less than 5 days gave inconsistent PCR results. B-globin and TCR-gamma products of up to 420 bp were amplified successfully from FFPE tissues, even after fixation for 72 h.

   Biospecimens

   • Tissue - Thyroid Gland
   • Tissue - Adrenal Gland
   • Tissue - Skin
   • Tissue - Bladder
   • Tissue - Colorectal
   • Tissue - Breast

   Preservative Types

   • Formalin
   • None (Fresh)

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	Electrophoresis
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Protein digestion	Proteinase k
   Analyte Extraction and Purification	Incubation duration/condition	1 day 2 days 3 days 4 days 5 days 6 days 7 days
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) None (fresh)
   Biospecimen Preservation	Time in fixative	3-72 h

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