DNA extraction from formalin-fixed laryngeal biopsies: Comparison of techniques

Acta Otolaryngol. 2011 Mar;131(3):330-3. doi: 10.3109/00016489.2010.528794. Epub 2010 Dec 10.

Abstract

Conclusion: PCR-quality DNA could be extracted from formalin-fixed paraffin-embedded (FFPE) samples with amplicons of at least 390 bp. Paraffin removal was not a necessary step. Proteinase K digestion was as efficient as the commercial kit for DNA extraction with a lower cost.

Objectives: To compare different DNA extraction protocols for FFPE samples and to describe the suitability of the extracted DNA for PCR reactions.

Methods: For deparaffinization the following techniques were compared: alkaline heat, xylene, and no removal. For DNA extraction, proteinase K digestion and organic extraction were compared. A commercial extraction kit was included as standard. DNA quality was assessed by PCR amplification of the HFE gene, for amplicons of 208 and 390 bp.

Results: Extraction with the commercial kit and proteinase K digestion were more efficient than other techniques, with no statistical difference between them for both amplicons. The proteinase K digestion buffer had a cost of U$ 0.2 per sample and the commercial kit of U$7 per sample.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biopsy
  • DNA, Neoplasm / genetics
  • DNA, Neoplasm / isolation & purification*
  • Formaldehyde / chemistry
  • Humans
  • Laryngeal Neoplasms / genetics*
  • Laryngeal Neoplasms / pathology
  • Nucleic Acid Amplification Techniques
  • Paraffin Embedding / methods*
  • Tissue Fixation / methods*

Substances

  • DNA, Neoplasm
  • Formaldehyde