Anna J Holmquist, Holly Tavris, Grace Kim, Lauren A Esposito, Brian L Fisher, Athena Lam
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引用次数: 0
摘要
自然历史藏品是生物多样性数据的宝贵宝库。大规模的基因组分析将极大地扩展博物馆藏品的实用性和可获取性,但基因组学方法的高成本和时间密集性限制了此类项目的开展,尤其是对无脊椎动物标本而言。本文介绍了一种创新、经济、高通量的方法,利用单相反向固定(SPRI)珠从不同的昆虫标本中提取基因组 DNA。我们优化了 PEG-8000 和 NaCl 的浓度,以平衡 DNA 的产量和纯度,将试剂成本降至每个样本 6-11 美分。我们的方法与三种广泛使用的提取方案进行了验证,DNA产量和扩增成功率与广泛使用的Qiagen DNeasy试剂盒相当。我们成功地以高通量的方式应用了该方法,从3786个昆虫标本中提取了DNA,这些标本涵盖了广泛的年龄、分类和组织类型。为了便于其他研究人员采用该方法,我们提供了一份详细的方案。通过改进任何分子项目中最关键的步骤之一,这种基于 SPRI 微珠的 DNA 提取方法具有实现大规模博物馆组学项目的巨大潜力,从而提高历史藏品在生物多样性研究和保护工作中的效用。
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 to extracting genomic DNA from diverse insect specimens using single-phase reverse immobilization (SPRI) beads. We optimized PEG-8000 and NaCl concentrations to balance DNA yield and purity, reducing reagent cost to 6-11 cents per sample. 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 3,786 insect specimens across a broad range of ages, taxonomies, and tissue types. A detailed protocol is 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.