19 – SAMPLE PREPARATION FROM PARAFFIN-EMBEDDED TISSUES
19 – SAMPLE PREPARATION FROM PARAFFIN-EMBEDDED TISSUES
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DOI：
10.1016/b978-0-12-372180-8.50023-8
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Deann K. Wright;M. Manos
中科院分区：
文献类型：
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作者：
Deann K. Wright;M. Manos
在分子水平上研究保存的组织的能力使得对大量患者进行回顾性研究成为可能，并且可以允许长期跟踪与疾病相关的遗传变化或传染源。保存人体组织的最常见方法是在福尔马林中固定，然后进行石蜡包埋。几个研究小组对从这些样本中提取的 DNA 进行了 Southern 印迹分析（Goelz 等人，1985 年；Dubeau 等人，1986 年）。虽然这种方法可以提供宝贵的信息，但它非常费力，因此不适合检查大量样品。来自旧样本或固定不当样本的 DNA 通常会被降解，无法通过 Southern 印迹法进行分析。使用 PCR 检查固定石蜡包埋组织中的 DNA 为检查大量样品提供了一种相对简单且极其灵敏的方法。（Impraim 等人，1987 年；Shibata 等人，1988a）此外，档案组织的 PCR 分析可以通过 5 至 10 μm 切片完成，而 Southern 印迹分析则需要更大量的组织。 PCR 不需要高分子量 DNA，因此可以分析可能不适合 Southern 印迹的变质样本
The ability to study preserved tissues at the molecular level makes possible retrospective studies on large numbers of patients and may permit the tracking, over long periods of time, of genetic changes or infectious agents that are associated with diseases. The most common method for preserving human tissue is fixation in formalin followed by paraffin embedding. Several research groups have performed Southern blot analyses on DNA extracted from such specimens (Goelz et al. 1985; Dubeau et al. 1986). While this approach can provide invaluable information, it is extremely laborious and therefore not suitable for the examination of large numbers of samples. DNA from old or improperly fixed samples is often degraded and cannot be analyzed by Southern blotting. The use of PCR to examine the DNA in fixed, paraffin-embedded tissues provides a relatively simple and extremely sensitive method for examining large numbers of samples.(Impraim et al. 1987; Shibata et al. 1988a) Additionally, PCR analysis of archival tissues can be accomplished with 5-to 10-μm sections, whereas Southern blot analysis requires larger amounts of tissue. PCR does not require high-molecular-weight DNA and therefore allows the analysis of deteriorated specimens that may be inappropriate for Southern blot