| 石蜡包埋组织DNA提取方法的比较及巢式PCR技术的应用 |
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| 引用本文： | Yan L,Sun B,Zhao X,Liu Z,Song W. 石蜡包埋组织DNA提取方法的比较及巢式PCR技术的应用[J]. 生物医学工程学杂志, 2011, 28(4): 795-799 |
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| 作者姓名： | Yan L Sun B Zhao X Liu Z Song W |
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| 作者单位： | 天津医科大学病理教研室; |
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| 基金项目： | 天津市科技发展基金资助项目(020127) |
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| 摘 要： | 确定从福尔马林固定、石蜡包埋(FFPE)组织中提取DNA的最佳方法,增强长片段PCR产物的成功扩增率,便于分子生物学与临床医学的有机结合。本文选取正常甲状腺FFPE标本20份,分别应用一步法、酚/氯仿抽提法和试剂盒法三种不同的方法提取总DNA,并应用传统PCR法和巢式PCR法对线粒体DNA(mtDNA)的D环区进行扩增。结果酚/氯仿抽提法所得样本DNA的OD260/D280比值最高,为1.703±0.086。一步法、酚/氯仿抽提法和试剂盒法提取DNA的普通PCR长片段扩增成功率分别为0%、5%和10%,而巢式PCR长片段扩增成功率分别为0%、95%和85%。可见用蛋白酶K消化、酚/氯仿纯化法提取的FFPE组织标本DNA质量可靠,巢式PCR技术是从石蜡标本中扩增长片段DNA的有效方法。
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| 关 键 词： | 石蜡包埋组织 DNA提取 巢式PCR 比较 |
Analysis of different methods of extracting DNA from paraffin-embedded tissues and the application of nest PCR |
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| Yan Limin,Sun Baocun,Zhao Xiulan,Liu Zenghui,Song Wenjing. Analysis of different methods of extracting DNA from paraffin-embedded tissues and the application of nest PCR[J]. Journal of biomedical engineering, 2011, 28(4): 795-799 |
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| Authors: | Yan Limin Sun Baocun Zhao Xiulan Liu Zenghui Song Wenjing |
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| Affiliation: | Yan Limin Sun Baocun Zhao Xiulan Liu Zenghui Song Wenjing(Department of Pathology,Tianjin Medical University,Tianjin 300070,China) |
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| Abstract: | The aim of this research was to explore the most optimal method of DNA extraction from formalin-fixed,paraffin-embedded(FFPE) tissues,and to improve the amplification of long fragments with the method.Three methods,one step method,phenol-chloroform extraction method,and genomic DNA purification kit method,were employed to extract DNA from twenty normal thyroid tissues which were fixed with formalin and embedded with paraffin.The highest proportionality of OD260/OD280 in the examples was obtained by phenol-c... |
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| Keywords: | Paraffin-embedded tissues DNA extraction Nest PCR Comparison |
| 本文献已被 CNKI PubMed 等数据库收录！ |
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