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在血点卡上的动物便物质的DNA提取协议

Ann-Katrin Llarena¹, Thomas H A Haverkamp², Wenche Støldal Gulliksen³

¹Department of Paraclinical Sciences, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Ås, Norway.

|May 12, 2025

概括

干燥防腐剂使大规模研究容易收集便样本. QIAsymphony® PowerFecal® Pro DNA Kit和2000万对结 (PE) 读数是动物便微生物组分析的最佳选择.

科学领域:

微生物学微生物学
基因组学就是基因组学.
生物信息学是一种生物信息学.

背景情况:

干燥防腐剂为大规模流行病学研究中收集便样本提供了一种成本效益高且在后勤方面简单的替代方案.
评估DNA提取方法对于从各种动物便 (犬,绵羊,马,牛,猪) 中获得高质量的DNA至关重要,这些便被保存在干血斑点卡上.
评估Illumina shotgun元基因组测序对于表征动物便微生物组的分类学和功能概况是必要的.

研究的目的:

确定一种高效的DNA提取方法,以从干血斑点卡上收集的动物便中提取高质量的DNA.
为了确定最优的测序深度,使用枪支元基因组学对便微生物组进行全面的分类学和功能分析.
评估Illumina shotgun metagenomic测序的适用性,用于表征动物便微生物组.

主要方法:

在猪便和模拟社区中测试了四种DNA提取套件:QIAsymphony® PowerFecal® Pro DNA Kit (两种方法),ZymoBIOMICSTM DNA Miniprep Kit,以及MagNA Pure 96 DNA和Viral NA小体积套件.
使用模拟社区的amplicon测序来控制提取偏差.
最有效的DNA提取方法是用于从各种动物获得的便样本的猎枪元基因组测序,读取深度各不相同.

主要成果:

这四种DNA提取方法都在模拟社区中产生了类似的细菌社区组成.
作为最佳方法,QIAsymphony® PowerFecal® Pro DNA Kit被选为最佳方法.
增加测序深度至500万对结 (PE) 读数增强了类丰富性,而功能分析的好处增加了超过2000万PE读数.

结论:

几种DNA提取方法适用于在血点卡上收集的便,QiaGEN在QiaSymphony平台上的血液和组织套件非常有效.
序列深度为2000万个PE读数,足以准确的分类学和共同的功能途径识别.
检测罕见的微生物特征需要比2000万个PE读数更高的测序努力.

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