Freshwater vertebrate metabarcoding on Illumina platforms using double-indexed primers of the mitochondrial 16S rRNA gene
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Data
2016-09-01
Autores
Vences, Miguel
Lyra, Mariana L. [UNESP]
Perl, R. G. Bina
Bletz, Molly C.
Stankovic, David
Lopes, Carla Martins [UNESP]
Jarek, Michael
Bhuju, Sabin
Geffers, Robert
Haddad, Celio F. B. [UNESP]
Título da Revista
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Título de Volume
Editor
Springer
Resumo
Metabarcoding is a promising tool for biodiversity inventories and other applications in conservation genetics. We developed a new pair of primers for efficient and affordable high-throughput analysis of a 250 base pair stretch of DNA from the mitochondrial 16S rRNA gene of vertebrates, especially amphibians and fishes. By adapting a double-indexed protocol for Illumina platforms, our approach allows pooling of hundreds of samples in a single sequencing run. We obtained high detection rates of 82-93 % for fish in two German streams, 70 % for mock mixes of DNA from amphibians and fishes, and could distinguish multiple gene copies in amphibians, probably caused by nuclear-mitochondrial transposed DNA or heteroplasmy.
Descrição
Palavras-chave
eDNA, Double-indexed primers, Amphibians, Discoglossus, NUMT
Como citar
Conservation Genetics Resources. Dordrecht: Springer, v. 8, n. 3, p. 323-327, 2016.