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Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays

Noninvasively collected samples are a common source of DNA in wildlife genetic studies. Currently, single nucleotide polymorphism (SNP) genotyping using microfluidic arrays is emerging as an easy-to-use and cost-effective methodology. Here we assessed the performance of microfluidic SNP arrays in ge...

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Autores principales: von Thaden, Alina, Cocchiararo, Berardino, Jarausch, Anne, Jüngling, Hannah, Karamanlidis, Alexandros A., Tiesmeyer, Annika, Nowak, Carsten, Muñoz-Fuentes, Violeta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589735/
https://www.ncbi.nlm.nih.gov/pubmed/28883428
http://dx.doi.org/10.1038/s41598-017-10647-w
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author von Thaden, Alina
Cocchiararo, Berardino
Jarausch, Anne
Jüngling, Hannah
Karamanlidis, Alexandros A.
Tiesmeyer, Annika
Nowak, Carsten
Muñoz-Fuentes, Violeta
author_facet von Thaden, Alina
Cocchiararo, Berardino
Jarausch, Anne
Jüngling, Hannah
Karamanlidis, Alexandros A.
Tiesmeyer, Annika
Nowak, Carsten
Muñoz-Fuentes, Violeta
author_sort von Thaden, Alina
collection PubMed
description Noninvasively collected samples are a common source of DNA in wildlife genetic studies. Currently, single nucleotide polymorphism (SNP) genotyping using microfluidic arrays is emerging as an easy-to-use and cost-effective methodology. Here we assessed the performance of microfluidic SNP arrays in genotyping noninvasive samples from grey wolves, European wildcats and brown bears, and we compared results with traditional microsatellite genotyping. We successfully SNP-genotyped 87%, 80% and 97% of the wolf, cat and bear samples, respectively. Genotype recovery was higher based on SNPs, while both marker types identified the same individuals and provided almost identical estimates of pairwise differentiation. We found that samples for which all SNP loci were scored had no disagreements across the three replicates (except one locus in a wolf sample). Thus, we argue that call rate (amplification success) can be used as a proxy for genotype quality, allowing the reduction of replication effort when call rate is high. Furthermore, we used cycle threshold values of real-time PCR to guide the choice of protocols for SNP amplification. Finally, we provide general guidelines for successful SNP genotyping of degraded DNA using microfluidic technology.
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spelling pubmed-55897352017-09-13 Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays von Thaden, Alina Cocchiararo, Berardino Jarausch, Anne Jüngling, Hannah Karamanlidis, Alexandros A. Tiesmeyer, Annika Nowak, Carsten Muñoz-Fuentes, Violeta Sci Rep Article Noninvasively collected samples are a common source of DNA in wildlife genetic studies. Currently, single nucleotide polymorphism (SNP) genotyping using microfluidic arrays is emerging as an easy-to-use and cost-effective methodology. Here we assessed the performance of microfluidic SNP arrays in genotyping noninvasive samples from grey wolves, European wildcats and brown bears, and we compared results with traditional microsatellite genotyping. We successfully SNP-genotyped 87%, 80% and 97% of the wolf, cat and bear samples, respectively. Genotype recovery was higher based on SNPs, while both marker types identified the same individuals and provided almost identical estimates of pairwise differentiation. We found that samples for which all SNP loci were scored had no disagreements across the three replicates (except one locus in a wolf sample). Thus, we argue that call rate (amplification success) can be used as a proxy for genotype quality, allowing the reduction of replication effort when call rate is high. Furthermore, we used cycle threshold values of real-time PCR to guide the choice of protocols for SNP amplification. Finally, we provide general guidelines for successful SNP genotyping of degraded DNA using microfluidic technology. Nature Publishing Group UK 2017-09-07 /pmc/articles/PMC5589735/ /pubmed/28883428 http://dx.doi.org/10.1038/s41598-017-10647-w Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
von Thaden, Alina
Cocchiararo, Berardino
Jarausch, Anne
Jüngling, Hannah
Karamanlidis, Alexandros A.
Tiesmeyer, Annika
Nowak, Carsten
Muñoz-Fuentes, Violeta
Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title_full Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title_fullStr Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title_full_unstemmed Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title_short Assessing SNP genotyping of noninvasively collected wildlife samples using microfluidic arrays
title_sort assessing snp genotyping of noninvasively collected wildlife samples using microfluidic arrays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589735/
https://www.ncbi.nlm.nih.gov/pubmed/28883428
http://dx.doi.org/10.1038/s41598-017-10647-w
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