档案尸体组织PCR分析。

S Bonin, F Petrera, B Niccolini, G Stanta
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引用次数: 133

摘要

背景:尸检来源的福尔马林固定组织和石蜡包埋组织是分子分析的重要来源,特别是在罕见疾病、神经病理学或分子流行病学研究中。由于DNA降解,只能从这种类型的组织中扩增出短序列,通常少于100个碱基。这就带来了问题,因为研究大基因的多态性和突变通常需要分析长序列。方法:开发一种简单的处理方法,以获得较长的DNA片段,用于分析档案死后石蜡包埋组织。结果:可以从死后组织中扩增出长达300个碱基的较长序列,而无需修改通常的DNA提取程序。为了获得更长的DNA片段,需要进行pcr前修复处理,通过填充单链断裂,然后进行剧烈的变性步骤。结论:这种简单处理的发展允许分析从档案死后石蜡包埋组织中获得的较长的DNA片段。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PCR analysis in archival postmortem tissues.

Background: Formalin fixed and paraffin wax embedded tissues of necropsy origin are an important source for molecular analysis especially in rare diseases, neuropathology, or molecular epidemiology studies. Because of DNA degradation, only short sequences can be amplified from this type of tissue, very often less than 100 bases. This poses problems because studies on polymorphism and mutations occurring in large genes often require the analysis of long sequences.

Methods: The development of a simple treatment to obtain longer fragments of DNA for the analysis of archival postmortem paraffin wax embedded tissues.

Results: It was possible to amplify longer sequences ranging up to 300 bases from postmortem tissues, with no modification to the usual DNA extraction procedures. To obtain longer stretches of DNA, a pre-PCR restoration treatment was required, by filling single strand breaks, followed by a vigorous denaturation step.

Conclusions: The development of this simple treatment allowed the analysis of longer fragments of DNA obtained from archival postmortem paraffin wax embedded tissues.

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