A simple and effective method to enrich endogenous DNA from mammalian faeces

被引:1
|
作者
Cui, Liang Yu [1 ,2 ]
Liu, Bo Yang [1 ,2 ]
Li, Hai Meng [3 ,4 ]
Zhu, Yi Xin [3 ,4 ]
Zhou, Yong Heng [1 ,2 ]
Su, Chang [1 ,2 ]
Tian, Yin Ping [1 ,2 ]
Xu, Hai Tao [5 ]
Liu, Dan [5 ]
Li, Xiao Ping [3 ,6 ]
Ma, Yue [1 ,6 ]
Jiang, Guang Shun [1 ,6 ,7 ]
Liu, Huan [3 ,6 ]
Yang, Shu Hui [1 ,6 ]
Lan, Tian Ming [3 ,6 ]
Xu, Yan Chun [1 ,2 ,6 ]
机构
[1] Northeast Forestry Univ, Coll Wildlife & Protected Area, Harbin, Peoples R China
[2] Natl Forestry & Grassland Adm Res Ctr Engn Techno, Harbin, Peoples R China
[3] BGI Shenzhen, State Key Lab Agr Genom, Shenzhen, Peoples R China
[4] Univ Chinese Acad Sci, Coll Life Sci, Beijing, Peoples R China
[5] Heilongjiang Siberian Tiger Pk, Harbin, Peoples R China
[6] Northeast Forestry Univ, BGI Life Sci Joint Res Ctr, Harbin, Peoples R China
[7] Natl Forestry & Grassland Adm Feline Res Ctr, Harbin, Peoples R China
基金
中国国家自然科学基金;
关键词
endogenous DNA; enrichment efficiency; genome resequencing; molecular marker; peri-extraction enrichment; scatology; MARTEN MARTES-MARTES; GENETIC-ANALYSIS; POPULATION-SIZE; STOOL; COLONOCYTES; EXTRACTION; SEPARATION; CAPTURE; GENOME; CELLS;
D O I
10.1111/1755-0998.13939
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页数:17
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