Efficiency evaluation of a DNA extraction and purification protocol on archival formalin-fixed and paraffin-embedded tissue
Efficiency evaluation of a DNA extraction and purification protocol on archival formalin-fixed and paraffin-embedded tissue
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DOI：
10.1016/j.forsciint.2009.09.004
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发表时间：
2010-01-30
影响因子：
2.2
通讯作者：
Ludes, B.
中科院分区：
文献类型：
--
作者：
Farrugia, A.;Keyser, C.;Ludes, B.
福尔马林固定石蜡包埋组织（FF-PET）是回顾性分子遗传学研究的宝贵资源，但从FF-PET中提取高质量的基因组DNA仍然是一个有问题的问题。尽管目前使用的DNA提取方法的范围，酚-氯仿提取和基于二氧化硅的纯化协议的关联，在古代DNA研究考古骨骼，据我们所知，尚未被用于从FF-PET的DNA提取。本研究通过定量PCR和多重扩增比较了从两种不同的FF-PET底物心脏和肝脏中提取和纯化DNA的三种方法的效率。我们表明，使用酚-氯仿和QIAamp DNA mini（R）试剂盒（Qiagen）的方法，结果表明，从肝组织中提取的DNA量比从肝组织中提取的DNA量更重要，从心脏中提取。常染色体STR分型的多重扩增得到部分等位基因的档案，只有小尺寸的产品（小于300个碱基）扩增，这表明从FF-PET提取的DNA degraded.In结论，这里提出的协议，以前描述的古骨骼的研究，应该找到应用在不同的分子研究涉及FF-PET材料。(C)2009爱思唯尔爱尔兰有限公司保留所有权利。
Formalin-fixed and paraffin-embedded tissue (FF-PET) is an invaluable resource for retrospective molecular genetic studies, but the extraction of high-quality genomic DNA from FF-PET is still a problematic issue. Despite the range of DNA extraction methods currently in use, the association of phenol-chloroform extraction and silica-based purification protocols, reported in ancient DNA studies on archaeological bones, has, to our knowledge, not been used for DNA extraction from FF-PET yet. The present study compared the efficiency of three DNA extraction and purification protocols from two different FF-PET substrates, heart and liver, by using quantitative PCR and multiplex amplification.We showed that the method, using phenol-chloroform and the QIAamp DNA mini(R) Kit (Qiagen), was the most effective DNA extraction and purification method and that the DNA quantity extracted from liver is statistically more important than that extracted from heart. Autosomal STR typing by multiplex amplifications gave partial allelic profiles with only small size products (less than 300 bases) amplified, suggesting that DNA extracted from FF-PET was degraded.In conclusion, the protocol presented here, previously described in studies on ancient bones, should find application in different molecular studies involving FF-PET material. (C) 2009 Elsevier Ireland Ltd. All rights reserved.