Effect of buffered formalin on amplification of DNA from paraffin wax embedded small biopsies using real-time PCR. Issue 6 (27th May 2004)

• Record Type: Journal Article
• Authors: Zsikla, V; Baumann, M; Cathomas, G
• Is Part Of: Journal of clinical pathology. Volume 57:Issue 6(2004)
• Page Start: 654
• Page End: 656
• Publication Date: 2004-05-27
• Subjects: unbuffered formalin fixation -- gastric and duodenal biopsy -- paraffin wax embedding -- real-time (TaqMan) polymerase chain reaction -- human β actin; PCR, polymerase chain reaction; Pathology -- Periodicals; Pathology, Molecular -- Periodicals; 616.0705
• DOI: 10.1136/jcp.2003.013961
• Languages: English
• ISSNs: 0021-9746

Abstract

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 β actin DNA extracted from upper gastrointestinal tract biopsies fixed in buffered and unbuffered formalin. Methods: Amounts of β 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 β actin DNA was significantly higher in biopsies fixed in buffered formalin than in those fixed in unbuffered formalin (median 2.8 × 10 4 and 5.3 × 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.