TY  - JOUR
A1  - Bayerl, Helmut
A1  - Kraus, Robert H. S.
A1  - Nowak, Carsten
A1  - Foerster, Daniel W.
A1  - Fickel, Jörns
A1  - Kühn, Ralph
T1  - Fast and cost-effective single nucleotide polymorphism (SNP) detection in the absence of a reference genome using semideep next-generation Random Amplicon Sequencing (RAMseq)
T2  - Molecular ecology resources
N2  - Biodiversity has suffered a dramatic global decline during the past decades, and monitoring tools are urgently needed providing data for the development and evaluation of conservation efforts both on a species and on a genetic level. However, in wild species, the assessment of genetic diversity is often hampered by the lack of suitable genetic markers. In this article, we present Random Amplicon Sequencing (RAMseq), a novel approach for fast and cost-effective detection of single nucleotide polymorphisms (SNPs) in nonmodel species by semideep sequencing of random amplicons. By applying RAMseq to the Eurasian otter (Lutra lutra), we identified 238 putative SNPs after quality filtering of all candidate loci and were able to validate 32 of 77 loci tested. In a second step, we evaluated the genotyping performance of these SNP loci in noninvasive samples, one of the most challenging genotyping applications, by comparing it with genotyping results of the same faecal samples at microsatellite markers. We compared (i) polymerase chain reaction (PCR) success rate, (ii) genotyping errors and (iii) Mendelian inheritance (population parameters). SNPs produced a significantly higher PCR success rate (75.5% vs. 65.1%) and lower mean allelic error rate (8.8% vs. 13.3%) than microsatellites, but showed a higher allelic dropout rate (29.7% vs. 19.8%). Genotyping results showed no deviations from Mendelian inheritance in any of the SNP loci. Hence, RAMseq appears to be a valuable tool for the detection of genetic markers in nonmodel species, which is a common challenge in conservation genetic studies.
KW  - high-throughput sequencing
KW  - Lutra lutra
KW  - nonmodel species
KW  - RAMseq
KW  - RAPD
KW  - variant detection
Y1  - 2018
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/54627
SN  - 1755-098X
SN  - 1755-0998
VL  - 18
IS  - 1
SP  - 107
EP  - 117
PB  - Wiley
CY  - Hoboken
ER  -