迈向大规模博物馆组学项目:一种从博物馆昆虫标本中获得历史DNA的高成本效益和高通量提取方法。

IF 5.5 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Anna Holmquist, Holly Tavris, Grace Kim, Lauren Esposito, Brian Fisher, Athena Lam
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引用次数: 0

摘要

自然历史收藏是生物多样性数据的宝贵宝库。大规模基因组分析将极大地扩大博物馆藏品的实用性和可及性,但基因组方法的高成本和时间密集的性质限制了这类项目,特别是对无脊椎动物标本。本文介绍了一种利用单相反向固定化(SPRI)微珠从多种昆虫标本中提取基因组DNA的创新、经济、高通量方法。我们优化了PEG-8000和NaCl浓度,以平衡DNA产率和纯度,将试剂成本降低到每个样品4.0-11.6美分,成本取决于样品类型。我们的方法与三种广泛使用的提取方案进行了验证,并显示出与广泛使用的Qiagen DNeasy试剂盒相当的DNA产量和扩增成功率。我们成功地以高通量的方式应用了该方案,从3786个昆虫标本中提取了DNA,这些标本跨越了广泛的年龄、分类和组织类型。提供了详细的协议和教学视频,以方便其他研究人员采用该方法。通过改进任何分子项目中最关键的步骤之一,这种基于SPRI珠的DNA提取方法具有实现大规模博物馆组学项目的巨大潜力,从而增加了生物多样性研究和保护工作的历史收藏品的效用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Towards Large-Scale Museomics Projects: A Cost-Effective and High-Throughput Extraction Method for Obtaining Historical DNA From Museum Insect Specimens.

Natural history collections serve as invaluable repositories of biodiversity data. Large-scale genomic analysis would greatly expand the utility and accessibility of museum collections, but the high cost and time-intensive nature of genomic methods limit such projects, particularly for invertebrate specimens. This paper presents an innovative, cost-effective and high-throughput approach for extracting genomic DNA from diverse insect specimens using single-phase reverse immobilisation (SPRI) beads. We optimised PEG-8000 and NaCl concentrations to balance DNA yield and purity, reducing reagent cost to 4.0-11.6¢ per sample, cost dependent on sample type. Our method was validated against three widely used extraction protocols and showed comparable DNA yield and amplification success to the widely used Qiagen DNeasy kit. We successfully applied the protocol in a high-throughput manner, extracting DNA from 3786 insect specimens across a broad range of ages, taxonomies and tissue types. A detailed protocol and instructional video are provided to facilitate the adoption of the method by other researchers. By improving one of the most crucial steps in any molecular project, this SPRI bead-based DNA extraction approach has significant potential for enabling large-scale museomics projects, thereby increasing the utility of historical collections for biodiversity research and conservation efforts.

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来源期刊
Molecular Ecology Resources
Molecular Ecology Resources 生物-进化生物学
CiteScore
15.60
自引率
5.20%
发文量
170
审稿时长
3 months
期刊介绍: Molecular Ecology Resources promotes the creation of comprehensive resources for the scientific community, encompassing computer programs, statistical and molecular advancements, and a diverse array of molecular tools. Serving as a conduit for disseminating these resources, the journal targets a broad audience of researchers in the fields of evolution, ecology, and conservation. Articles in Molecular Ecology Resources are crafted to support investigations tackling significant questions within these disciplines. In addition to original resource articles, Molecular Ecology Resources features Reviews, Opinions, and Comments relevant to the field. The journal also periodically releases Special Issues focusing on resource development within specific areas.
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