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从存档福尔马林固定石蜡包埋尸检组织中回收核酸的方法比较。

Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues.

机构信息

McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, Ontario L8S4L9, Canada.

出版信息

Anal Biochem. 2010 May 1;400(1):110-7. doi: 10.1016/j.ab.2010.01.014. Epub 2010 Jan 15.

Abstract

Archival formalin-fixed paraffin-embedded (FFPE) human tissue collections are typically in poor states of storage across the developing world. With advances in biomolecular techniques, these extraordinary and virtually untapped resources have become an essential part of retrospective epidemiological studies. To successfully use such tissues in genomic studies, scientists require high nucleic acid yields and purity. In spite of the increasing number of FFPE tissue kits available, few studies have analyzed their applicability in recovering high-quality nucleic acids from archived human autopsy samples. Here we provide a study involving 10 major extraction methods used to isolate total nucleic acid from FFPE tissues ranging in age from 3 to 13years. Although all 10 methods recovered quantifiable amounts of DNA, only 6 recovered quantifiable RNA, varying considerably and generally yielding lower DNA concentrations. Overall, we show quantitatively that TrimGen's WaxFree method and our in-house phenol-chloroform extraction method recovered the highest yields of amplifiable DNA, with considerable polymerase chain reaction (PCR) inhibition, whereas Ambion's RecoverAll method recovered the most amplifiable RNA.

摘要

存档的福尔马林固定石蜡包埋(FFPE)人体组织在发展中国家的储存状况通常较差。随着生物分子技术的进步,这些非凡且几乎未开发的资源已成为回顾性流行病学研究的重要组成部分。为了在基因组研究中成功使用这些组织,科学家需要高核酸产量和纯度。尽管 FFPE 组织试剂盒的数量不断增加,但很少有研究分析它们在从存档的人体尸检样本中提取高质量核酸方面的适用性。在这里,我们提供了一项涉及 10 种主要提取方法的研究,这些方法用于从年龄在 3 至 13 岁的 FFPE 组织中分离总核酸。尽管所有 10 种方法都回收了可定量的 DNA,但只有 6 种方法回收了可定量的 RNA,差异很大,通常 DNA 浓度较低。总的来说,我们定量地表明,TrimGen 的 WaxFree 方法和我们内部的酚氯仿提取方法回收了最高产量的可扩增 DNA,但聚合酶链反应(PCR)抑制相当严重,而 Ambion 的 RecoverAll 方法回收了最多的可扩增 RNA。

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