Liang Yu Cui, Bo Yang Liu, Hai Meng Li, Yi Xin Zhu, Yong Heng Zhou, Chang Su, Yin Ping Tian, Hai Tao Xu, Dan Liu, Xiao Ping Li, Yue Ma, Guang Shun Jiang, Huan Liu, Shu Hui Yang, Tian Ming Lan, Yan Chun Xu
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引用次数: 0
Abstract
Utilization of faeces has long been a popular approach for genetic and ecological studies of wildlife. However, the success of molecular marker genotyping and genome resequencing is often unpredictable due to insufficient enrichment of endogenous DNA in the total faecal DNA that is dominated by bacterial DNA. Here, we report a simple and cheap method named PEERS to predominantly lyse animal cells over bacteria by using sodium dodecyl sulphate so as to discharge endogenous DNA into liquid phase before bacterial DNA. By brief centrifugation, total DNA with enriched endogenous fraction can be extracted from the supernatant using routine methods. Our assessments showed that the endogenous DNA extracted by PEERS was significantly enriched for various types of faeces from different species, preservation time and conditions. It significantly improves the genotyping correctness and efficiency of genome resequencing with the total additional cost of $ 0.1 and a short incubation step to treat a faecal sample. We also provide methods to assess the enrichment efficiency of mitochondrial and nuclear DNA and models to predict the usability of faecal DNA for genotyping of short tandem repeat, single-nucleotide polymorphism and whole-genome resequencing.
长期以来,利用粪便进行野生动物遗传和生态研究一直是一种流行的方法。然而,分子标记基因分型和基因组重测序的成功与否往往难以预测,原因是在以细菌 DNA 为主的总粪便 DNA 中,内源 DNA 的富集程度不够。在此,我们报告了一种名为 PEERS 的简单而廉价的方法,该方法利用十二烷基硫酸钠主要裂解动物细胞而非细菌,从而在细菌 DNA 之前将内源 DNA 排入液相。通过简短的离心,可以用常规方法从上清液中提取出富含内源部分的总 DNA。我们的评估结果表明,对于不同种类、保存时间和条件的各种粪便,PEERS 提取的内源性 DNA 都有明显的富集。它大大提高了基因分型的正确性和基因组重测序的效率,而处理粪便样本的额外总成本仅为 0.1 美元,培养步骤也很短。我们还提供了评估线粒体和核 DNA 富集效率的方法,以及预测粪便 DNA 在短串联重复、单核苷酸多态性和全基因组重测序基因分型中可用性的模型。
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.