Non-destructive DNA extraction from specimens and environmental samples using DESS preservation solution for DNA barcoding

Eri Ogiso-Tanaka, Minako Abe Ito, Daisuke Shimada
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Abstract

DESS is a widely used storage solution for the preservation of DNA from biological tissue samples. DESS comprises 20% dimethyl sulfoxide, 250 mM ethylenediaminetetraacetic acid, and saturated sodium chloride, and its efficacy has been confirmed in various taxa and tissues. DESS enables the stable, long-term preservation of both sample morphology and DNA. However, to access the DNA, excising a portion of the sample was necessary. Although DNA is a valuable source of information for species identification, DNA extraction can result in the loss of an entire sample or segment, especially in small-sized organisms, thereby compromising specimen value. Therefore, establishing non-destructive DNA extraction techniques is imperative. Thus, this paper presents a protocol for conducting non-destructive DNA extraction and DNA barcoding using a portion of the DESS supernatant obtained from a nematode specimen. This method was successfully employed for DNA barcoding of nematodes that were stored in DESS at room temperature (-10~35˚C) for ten years. Moreover, the method can be potentially applied in the preservation and non-destructive extraction of DNA from specimens of various species. Following sample collection, a bulk environmental sample from sediment and seagrass is immediately immersed in DESS in the field. Subsequently, DNA is extracted from the supernatant solution, allowing non-destructive DNA barcoding. Overall, this paper presents comprehensive protocols for DNA extraction from DESS supernatants and demonstrates their practical application using meiofauna (small animals) and diatoms as examples.
使用 DESS 保存解决方案从标本和环境样本中无损提取 DNA,用于 DNA 条形码分析
DESS 是一种广泛使用的生物组织样本 DNA 保存液。DESS 由 20% 二甲基亚砜、250 mM 乙二胺四乙酸和饱和氯化钠组成,其功效已在不同类群和组织中得到证实。DESS 可以长期稳定地保存样本形态和 DNA。不过,要获取 DNA,必须切除部分样本。虽然 DNA 是物种鉴定的宝贵信息来源,但 DNA 提取可能会导致整个样本或样本片段的损失,尤其是在小型生物体中,从而影响标本的价值。因此,建立非破坏性的 DNA 提取技术势在必行。因此,本文介绍了一种利用从线虫标本中获得的部分 DESS 上清液进行非破坏性 DNA 提取和 DNA 条形码的方法。该方法被成功地用于对在室温(-10~35˚C)下储存在 DESS 中的线虫进行 DNA 条形编码。此外,该方法还可用于保存和无损提取不同物种标本中的 DNA。样本采集完成后,在野外立即将沉积物和海草的大量环境样本浸入 DESS 中。随后,从上清液中提取 DNA,从而实现非破坏性 DNA 条形编码。总之,本文介绍了从 DESS 上清液中提取 DNA 的综合方案,并以小型动物和硅藻为例展示了其实际应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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