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评估替代环境毒物福尔马林的方法,以保存生物标本中的 DNA。

Assessment of alternatives to environmental toxic formalin for DNA conservation in biological specimens.

机构信息

INSERM-UMR1037, Université de Toulouse, Centre de Recherches en Cancérologie de Toulouse (CRCT), Equipe Labellisée Ligue Contre le Cancer, Laboratoire d'Excellence Toulouse Cancer (LabEx TOUCAN), 31037, Toulouse, France.

Muséum d'Histoire Naturelle de Toulouse, 35 allées Jules Guesde, 31000, Toulouse, France.

出版信息

Environ Sci Pollut Res Int. 2017 Jul;24(20):16985-16993. doi: 10.1007/s11356-017-9349-y. Epub 2017 Jun 3.

Abstract

One essential step of museum and clinical specimen preservation is immersion in a fixative fluid to prevent degradation. Formalin is the most largely used fixative, but its benefit is balanced with its toxic and carcinogenic status. Moreover, because formalin-fixation impairs nucleic acids recovery and quality, current museum wet collections and formalin-fixed, paraffin-embedded clinical samples do not represent optimal tanks of molecular information. Our study has been developed to compare formalin to two alternative fixatives (RCL2® and ethanol) in a context of molecular exploitation. Based on a unique protocol, we created mammalian fixed collections, simulated the impact of time on preservation using an artificial ageing treatment and followed the evolution of specimens' DNA quality. DNA extraction yield, purity, visual integrity and qualitative and quantitative ability to amplify the Cox1 gene were assessed. Our results show that both RCL2 and ethanol exhibit better performances than formalin. They do not impair DNA extraction yield, and more importantly, DNA alteration is delayed over the preservation step. The use of RCL2 or ethanol as fixative in biological collections may insure a better exploitation of the genetic resources they propose.

摘要

博物馆和临床标本保存的一个重要步骤是将其浸泡在固定液中以防止降解。福尔马林是最广泛使用的固定剂,但它的好处与其毒性和致癌性相平衡。此外,由于福尔马林固定会损害核酸的回收和质量,因此当前的博物馆湿标本和福尔马林固定、石蜡包埋的临床标本不能代表最佳的分子信息库。我们的研究旨在比较福尔马林与两种替代固定剂(RCL2®和乙醇)在分子利用方面的情况。基于独特的方案,我们创建了哺乳动物固定标本集,模拟了时间对保存的影响,使用人工老化处理,并跟踪标本 DNA 质量的演变。评估了 DNA 提取产量、纯度、视觉完整性以及扩增 Cox1 基因的定性和定量能力。我们的结果表明,RCL2 和乙醇的性能均优于福尔马林。它们不会损害 DNA 提取产量,更重要的是,在保存步骤中 DNA 改变被延迟。在生物标本集中使用 RCL2 或乙醇作为固定剂可以确保更好地利用它们提供的遗传资源。

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