An alternative protocol for DNA extraction from formalin fixed and paraffin wax embedded tissue
An alternative protocol for DNA extraction from formalin fixed and paraffin wax embedded tissue
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DOI：
10.1136/jcp.2004.021352
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发表时间：
2005-08-01
影响因子：
3.4
通讯作者：
Ashton-Prolla, P
中科院分区：
文献类型：
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作者：
Coura, R;Prolla, JC;Ashton-Prolla, P
背景：从石蜡包埋的组织中提取 DNA 需要特殊的方案，大多数描述的方法报告扩增成功率为 60 - 80%。目的：提出一种简单且廉价的方案，包括二甲苯/乙醇脱蜡，然后是基于试剂盒的提取。方法：二甲苯/乙醇脱蜡后进行长时间的再水化步骤和基于试剂盒的 DNA 提取步骤。结果：该方法对 121 至 227 的片段产生 100% 的扩增成功率结论：这种经济有效且不费力的方案可以成功地从填塞福尔马林固定石蜡包埋组织中提取 DNA，并有助于在回顾性研究中对大量档案标本进行分子分析。
Background: DNA extraction from paraffin wax embedded tissue requires special protocols, and most described methods report an amplification success rate of 60 - 80%.Aims: To propose a simple and inexpensive protocol consisting of xylene/ ethanol dewaxing, followed by a kit based extraction.Method: Xylene/ ethanol dewaxing was followed by a long rehydration step and a kit based DNA extraction step.Results: This method produced a 100% amplification success rate for fragments of 121 to 227 bp for tamponated formalin fixed paraffin wax embedded tissue.Conclusion: This cost effective and non- laborious protocol can successfully extract DNA from tamponated formalin fixed paraffin wax embedded tissue and should facilitate the molecular analysis of a large number of archival specimens in retrospective studies.