从Bouin固定组织中获取的DNA和RNA。
DNA and RNA obtained from Bouin's fixed tissues.
作者信息
Bonin S, Petrera F, Rosai J, Stanta G
机构信息
Department of Clinical, Morphological and Technological Sciences, University of Trieste, Cattinara Hospital, 34149 Trieste, Italy.
出版信息
J Clin Pathol. 2005 Mar;58(3):313-6. doi: 10.1136/jcp.2004.016477.
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.
背景
在许多国家，诸如Bouin氏液之类的酸性固定剂的使用限制了存档组织用于分子分析。酸性环境是DNA降解的主要原因之一。此外，在这类固定组织中提取RNA很困难。
目的
从Bouin氏固定组织中扩增DNA和RNA。
方法
从20例常规用Bouin氏固定液固定的乳腺癌样本中提取DNA和RNA。使用聚合酶链反应（PCR）对DNA（有或没有修复）以及对RNA进行逆转录PCR，使用产生不同长度扩增子的引物对几个基因进行扩增。
结果
Bouin氏固定液的酸性环境损害了DNA和RNA。然而，当RNA的扩增子长度减少到约80 bp且DNA的扩增子长度减少到100 - 200 bp时，扩增成功，特别是如果进行了DNA重建程序。
结论
从Bouin氏固定并石蜡包埋的组织中回收和分析DNA和RNA是可能的。
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