Effect of buffered formalin on amplification of DNA from paraffin wax embedded small biopsies using real-time PCR

J Clin Pathol. 2004 Jun;57(6):654-6. doi: 10.1136/jcp.2003.013961.

Abstract

Background: The isolation of good quality DNA from routinely fixed and processed biopsy samples is crucial for the success of subsequent molecular analysis.

Aims: To compare the amount of beta actin DNA extracted from upper gastrointestinal tract biopsies fixed in buffered and unbuffered formalin.

Methods: Amounts of beta actin DNA extracted from forceps biopsies of the upper gastrointestinal tract fixed in unbuffered (n = 22) and buffered formalin (n = 16) were estimated by quantitative real-time polymerase chain reaction.

Results: The yield of beta actin DNA was significantly higher in biopsies fixed in buffered formalin than in those fixed in unbuffered formalin (median 2.8 x 10(4) and 5.3 x 10(2) DNA molecules, respectively; p < 0.005). Furthermore, fixation in buffered formalin led to a more reproducible DNA extraction, as indicated by the coefficient of variation (1.0 and 2.2, respectively).

Conclusions: This study indicates that tissue samples should be fixed in buffered formalin to facilitate the use of molecular pathology analysis in routine biopsy material.

MeSH terms

  • Actins / genetics*
  • Biopsy
  • Buffers
  • DNA / analysis*
  • Digestive System / chemistry
  • Formaldehyde
  • Humans
  • Paraffin Embedding
  • Polymerase Chain Reaction / methods*
  • Tissue Fixation / methods

Substances

  • Actins
  • Buffers
  • Formaldehyde
  • DNA