一种从福尔马林固定石蜡包埋组织中提取DNA的替代方案。
An alternative protocol for DNA extraction from formalin fixed and paraffin wax embedded tissue.
作者信息
Coura R, Prolla J C, Meurer L, Ashton-Prolla P
机构信息
Genetics and Molecular Biology Post-graduation Program, Federal University from Rio Grande do Sul, Hospital de Clínicas de Porto Alegre, Brazil, CEP 90035-003.
出版信息
J Clin Pathol. 2005 Aug;58(8):894-5. doi: 10.1136/jcp.2004.021352.
Abstract
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.
摘要
背景
从石蜡包埋组织中提取DNA需要特殊方案，大多数已描述的方法报告的扩增成功率为60%-80%。
目的
提出一种简单且成本低廉的方案，包括二甲苯/乙醇脱蜡，随后进行基于试剂盒的提取。
方法
二甲苯/乙醇脱蜡后进行长时间复水步骤和基于试剂盒的DNA提取步骤。
结果
对于用棉塞处理的福尔马林固定石蜡包埋组织，该方法对121至227 bp片段的扩增成功率为100%。
结论
这种经济高效且不费力的方案能够成功地从用棉塞处理的福尔马林固定石蜡包埋组织中提取DNA，并且应有助于回顾性研究中大量存档标本的分子分析。
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