Development of a Noninvasive Genotyping‐In‐Thousands (GTseq) Panel for Long‐Term Conservation of Western Great Lakes Gray Wolves (Canis lupus) DOI Creative Commons
Samuel D. Hervey, Bridgett M. vonHoldt, Mark C. Romanski

et al.

Ecology and Evolution, Journal Year: 2025, Volume and Issue: 15(4)

Published: April 1, 2025

ABSTRACT The application of noninvasive genetic methods toward the field conservation has increased our understanding many wildlife populations that are difficult to sample, allowing for better management. In molecular ecology, use sampling became widely feasible with advent microsatellites, a highly polymorphic, short‐length marker could be genotyped from low‐quality DNA sources. Despite decades use, microsatellite panels continue suffer high genotyping error rates, allelic dropout, and limited reproducibility across laboratories. To address these issues, single nucleotide polymorphisms (SNPs) offer advantages such as lower avoidance dropout due consistent allele length, automated calling through bioinformatic pipelines, reducing human subjectivity error. Given SNPs provide relative microsatellites marker, SNP specifically, method genotyping‐in‐thousands by sequencing (GTseq) gained popularity. Here, we developed GTseq panel western Great Lakes canids comprised 196 loci, capable species identification, accurately inferring sex (97.2%), identifying unique individuals (probability identity = 6.71e −41 ), assigning relationships (false positive rate 9.34e −14 genotypes low (0.39%). an attempt improve success samples, found while increasing number PCR cycles yielded higher percentage it also on‐target reads in negative controls. We suggest approaching this manipulation caution emphasize importance including reporting Further, quantitative was powerful estimate host‐specific concentrations, enabling conservative sample selection library preparation respect affordability.

Language: Английский

Development of a Noninvasive Genotyping‐In‐Thousands (GTseq) Panel for Long‐Term Conservation of Western Great Lakes Gray Wolves (Canis lupus) DOI Creative Commons
Samuel D. Hervey, Bridgett M. vonHoldt, Mark C. Romanski

et al.

Ecology and Evolution, Journal Year: 2025, Volume and Issue: 15(4)

Published: April 1, 2025

ABSTRACT The application of noninvasive genetic methods toward the field conservation has increased our understanding many wildlife populations that are difficult to sample, allowing for better management. In molecular ecology, use sampling became widely feasible with advent microsatellites, a highly polymorphic, short‐length marker could be genotyped from low‐quality DNA sources. Despite decades use, microsatellite panels continue suffer high genotyping error rates, allelic dropout, and limited reproducibility across laboratories. To address these issues, single nucleotide polymorphisms (SNPs) offer advantages such as lower avoidance dropout due consistent allele length, automated calling through bioinformatic pipelines, reducing human subjectivity error. Given SNPs provide relative microsatellites marker, SNP specifically, method genotyping‐in‐thousands by sequencing (GTseq) gained popularity. Here, we developed GTseq panel western Great Lakes canids comprised 196 loci, capable species identification, accurately inferring sex (97.2%), identifying unique individuals (probability identity = 6.71e −41 ), assigning relationships (false positive rate 9.34e −14 genotypes low (0.39%). an attempt improve success samples, found while increasing number PCR cycles yielded higher percentage it also on‐target reads in negative controls. We suggest approaching this manipulation caution emphasize importance including reporting Further, quantitative was powerful estimate host‐specific concentrations, enabling conservative sample selection library preparation respect affordability.

Language: Английский

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