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Genomic-scale capture and sequencing of endogenous DNA from feces.
Mol Ecol. 2010 Nov 3;
Authors: Perry GH, Marioni JC, Melsted P, Gilad Y
Genomic-level analyses of DNA from non-invasive sources would facilitate powerful conservation and evolutionary studies in natural populations of endangered and otherwise elusive species. However, the typical low quantity and poor quality of DNA that is extracted from non-invasive samples have generally precluded such work. Here we apply a modified DNA capture protocol that, when used in combination with massively-parallel sequencing technology, facilitates efficient and highly-accurate resequencing of megabases of specified nuclear genomic regions from fecal DNA samples. We validated our approach by comparing genetic variants identified from corresponding fecal and blood DNA samples of six western chimpanzees (Pan troglodytes verus) across more than 1.5 megabases of chromosome 21, chromosome X, and the complete mitochondrial genome. Our results suggest that it is now feasible to conduct genomic studies in natural populations for which constraints on invasive sampling have otherwise long been a barrier. The data we collected also provided an opportunity to examine western chimpanzee genetic diversity at unprecedented scale. Despite high mitochondrial genome diversity (? = 0.585%), western chimpanzees have a low ratio (0.42) of X chromosomal (? = 0.034%) to autosomal (chromosome 21 ? = 0.081%) sequence diversity, a pattern that may reflect an unusual demographic history of this subspecies.
PMID: 21054605 [PubMed - as supplied by publisher]
More...
Genomic-scale capture and sequencing of endogenous DNA from feces.
Mol Ecol. 2010 Nov 3;
Authors: Perry GH, Marioni JC, Melsted P, Gilad Y
Genomic-level analyses of DNA from non-invasive sources would facilitate powerful conservation and evolutionary studies in natural populations of endangered and otherwise elusive species. However, the typical low quantity and poor quality of DNA that is extracted from non-invasive samples have generally precluded such work. Here we apply a modified DNA capture protocol that, when used in combination with massively-parallel sequencing technology, facilitates efficient and highly-accurate resequencing of megabases of specified nuclear genomic regions from fecal DNA samples. We validated our approach by comparing genetic variants identified from corresponding fecal and blood DNA samples of six western chimpanzees (Pan troglodytes verus) across more than 1.5 megabases of chromosome 21, chromosome X, and the complete mitochondrial genome. Our results suggest that it is now feasible to conduct genomic studies in natural populations for which constraints on invasive sampling have otherwise long been a barrier. The data we collected also provided an opportunity to examine western chimpanzee genetic diversity at unprecedented scale. Despite high mitochondrial genome diversity (? = 0.585%), western chimpanzees have a low ratio (0.42) of X chromosomal (? = 0.034%) to autosomal (chromosome 21 ? = 0.081%) sequence diversity, a pattern that may reflect an unusual demographic history of this subspecies.
PMID: 21054605 [PubMed - as supplied by publisher]
More...