Details:

Journal Title:
Conservation Genetics Resources
Personal Author:
Gruenthal, K.M. ; Larson, W.A.
Gruenthal, K.M. ; Larson, W.A. Less -
NOAA Program & Office:
NMFS (National Marine Fisheries Service) ; AFSC (Alaska Fisheries Science Center)
NMFS (National Marine Fisheries Service) ; AFSC (Alaska Fisheries Science Center) Less -
Description:
Microsatellites have been a staple of population genetics research for over three decades, and many large datasets have been generated with these markers. Microsatellites have been used, for example, to conduct genetic monitoring and construct large multigeneration pedigrees as well as genotype thousands of individuals from a given species to create high-resolution baselines of spatial genetic structure. However, the capillary electrophoresis (CE) approach used to genotype microsatellites is inefficient compared to newer genotyping-by-sequencing (GBS) approaches, and researchers have begun transitioning away from CE. Backward compatibility between GBS and CE would facilitate a seamless transition to a more efficient chemistry, while ensuring that research based on CE panels could continue. Here, we explore the feasibility of converting a legacy panel of 15 microsatellites developed for muskellunge (Esox masquinongy) from CE to GBS chemistry. Muskellunge are an important sportfish in the Great Lakes region, and the existing microsatellite panel has been used to genotype thousands of samples to develop a region-wide baseline of genetic structure. We successfully converted all 15 microsatellites to GBS chemistry. GBS produced high genotyping rates (98%) and had high concordance with CE microsatellite genotypes (99%). Conversion to GBS required redesign of some primers and pairs to shorten amplicon length and adjust melting temperatures, optimization of primer concentrations, and comparisons with CE genotypes to optimize GBS genotyping parameters; however, none of these steps were especially onerous. Our results demonstrate that it is highly feasible to convert legacy CE panels to GBS, ensuring seamless continuation of important, often long-term research.
Keywords:
[+]
Capillary Electrophoresis Genetics Microsatellites (Genetics) Muskellunge Population Genetics Esox Masquinongy Genotyping-by-sequencing (GBS)
Source:
Conservation Genetics Resources (2021) 13:151–159
DOI:
https://doi.org/10.1007/s12686-020-01185-1
Format:
PDF
Document Type:
Journal Article
Rights Information:
Accepted Manuscript
Compliance:
Submitted
Main Document Checksum:
[+]
urn:sha256:0a0b99e2de578e8505dc49c0428430e631a01723c2534a6e076000d8bc36b1d0
Download URL:
https://repository.library.noaa.gov/view/noaa/52224/noaa_52224_DS1.pdf
File Type:

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