DNA and RNA obtained from Bouin’s fixed tissues
- 1ICGEB-International Centre for Genetic Engineering and Biotechnology, 99 Padriciano, 34012 Trieste, Italy
- 2Department of Clinical, Morphological and Technological Sciences, University of Trieste, Cattinara Hospital, 447 Strada di Fiume, 34149 Trieste, Italy
- 3Department of Pathology, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milano, Italy
- Correspondence to: Dr G Stanta International Centre for Genetic Engineering and Biotechnology, 99 Padriciano, 34012 Trieste, Italy; stantaicgeb.org
- Accepted 3 August 2004
Abstract
Background: The use in many countries of acid fixatives, such as Bouin’s solution, has limited the use of archival tissue for molecular analysis. An acidic environment is one of the main causes of DNA degradation. Moreover, RNA extraction is difficult in these types of fixed tissues.
Aims: To amplify DNA and RNA from Bouin’s fixed tissues.
Methods: DNA and RNA were extracted from 20 breast cancer samples that had been routinely fixed in Bouin’s fixative. Amplification of several genes using primers that produced amplicons of different lengths was carried out using the polymerase chain reaction (PCR) for DNA (with and without restoration) and reverse transcription PCR for RNA.
Results: The acid environment of Bouin’s fixative damaged both DNA and RNA. However, amplification was successful when the amplicon length was reduced to about 80 bp for RNA and 100–200 bp for DNA, especially if submitted to DNA reconstruction procedures.
Conclusions: It is possible to recover and analyse DNA and RNA from Bouin’s fixed and paraffin wax embedded tissues.
- EGFR, epidermal growth factor receptor
- GAPDH, human glyceraldehyde-3-phosphate dehydrogenase
- PCR, polymerase chain reaction
- RT, reverse transcription
- TTR, transthyretin gene
Footnotes
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The first two authors contributed equally to this study
