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. 2005 Mar;58(3):313-6.
doi: 10.1136/jcp.2004.016477.

DNA and RNA obtained from Bouin's fixed tissues

S Bonin  1 , F Petrera, J Rosai, G Stanta
Affiliations

DNA and RNA obtained from Bouin's fixed tissues

S Bonin et al. J Clin Pathol. 2005 Mar.

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.

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Figures

Figure 1
Figure 1
PCR products for TTR1 and TTR2 analysis in Bouin’s solution fixed, paraffin wax embedded tissues. (A) TTR1 DNA (291 bp) with the restoration step. Lane 1, molecular size marker; lanes 2–10, DNA from breast cancer cases. (B) TTR1 DNA (291 bp) with the restoration step. Lanes 1–11, DNA from breast cancer cases. (C) TTR2 gene DNA (333 bp) with the restoration step. Lane 1, molecular size marker; lanes 2–10, DNA from breast cancer cases. (D) TTR gene DNA (333 bp) with the restoration step. Lanes 1–11, DNA from breast cancer cases. PCR, polymerase chain reaction; TTR, transthyretin gene.
Figure 2
Figure 2
RT-PCR products in Bouin’s solution fixed, paraffin wax embedded tissues. (A) mRNA analysis for the β actin gene (100 bp). Lane 1, molecular size marker; lanes 2–10, RNA obtained from breast cancer cases. (B) mRNA analysis for β actin gene (100 bp). Lanes 1–11, RNA obtained from breast cancer cases. (C) mRNA analysis for β actin gene (120 bp). Lane 1, molecular size marker; lanes 2–10, RNA obtained breast cancer cases. (D) mRNA analysis for β actin gene (120 bp). Lanes 1–11, RNA obtained from breast cancer cases. RT-PCR, reverse transcription polymerase chain reaction.
See this image and copyright information in PMC

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