I suggest you to contact Kristina von Rintelen at Naturkundemuseum in Berlin

Dear .... If this file usful for you ...you can contact the Authors in paper 

Freshwater vertebrate metabarcoding on Illumina platforms using

double-indexed primers of the mitochondrial 16S rRNA gene

Miguel Vences1 • Mariana L. Lyra2 • R. G. Bina Perl1 • Molly C. Bletz1 •

David Stankovic´

3 • Carla Martins Lopes2 • Michael Jarek4 • Sabin Bhuju4 •

Robert Geffers4 • Ce´lio F. B. Haddad2 • Sebastian Steinfartz1

Received: 10 February 2016 / Accepted: 21 May 2016 / Published online: 28 May 2016

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Springer Science+Business Media Dordrecht 2016

Abstract Metabarcoding is a promising tool for biodi-

versity 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.

Keywords eDNA -

Double-indexed primers -

Amphibians -

Discoglossus -

NUMT

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