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福尔马林固定石蜡包埋组织3种DNA提取方法比较
Comparison of three methods for extraction of DNA from formalin fixed paraffin-embedded tissues
【摘要】 目的探讨经福尔马林固定1d石蜡包埋组织(FFPET)提取DNA的简易有效方法。方法比较水浴加热、微波加热和二甲苯脱蜡的效果。组织脱蜡后分别采用Chelex-100+层析柱纯化法、DNA IQTM试剂盒磁珠提取法和Chelex-100+磁珠纯化法提取DNA;实时荧光定量PCR技术定量DNA;荧光标记毛细管电泳技术进行STR分型。结果二甲苯脱蜡的效果好于其他两种加热的脱蜡方法(P<0.05)。Chelex-100+层析柱纯化所获得的DNA量显著高于其他两种方法(P<0.05)。结论二甲苯脱蜡、Chelex-100+层析柱纯化法是一种简单、有效的FFPET处理方法。
【Abstract】 Objective To explore an optimal method for extraction of DNA from formalin fixed paraffin-embedded tissues.Methods Compare the effect of the water bath heating,microwave heating,and xylene dewaxing.The specimens of each group were treated by Chelex-100 combined with QIAquick PCR Purification Kit,DNA IQTM kit,Chelex-100 combined with DNA IQTM kit,respectively.The quality of obtained DNA was analyzed by PCR quantification.DNA was amplified with sinofiler PCR Amplification kit.Results The yield of DNA dewaxed by xylene and extracted by Chelex-100 combined with purified by QIAquick PCR Purification Kit is significantly higher than those treated by other methods.Conclusion The Chelex-100 extraction combined with QIAquick PCR Kit purification is very suitable for extracting DNA from formalin fixed paraffin-embedded tissues.
【Key words】 forensic biological evidence; formalin fixed; paraffin-embedded tissues; DNA quantification; STR typing;
- 【文献出处】 中国法医学杂志 ,Chinese Journal of Forensic Medicine , 编辑部邮箱 ,2013年03期
- 【分类号】D919
- 【被引频次】5
- 【下载频次】413