Alkaline dip DNA extraction from skin mucus for high-throughput sexing of sterlets (Acipenser ruthenus)

Studies on simple DNA extraction methods from fish mucus, which is an ideal resource for noninvasive sampling, are scarce. In the aquaculture of sturgeons such as sterlets (Acipenser ruthenus), a high-throughput genetic sexing method is needed, as only females are reared to maturity for their roe. H...

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Détails bibliographiques
Publié dans:BioTechniques. - 1991. - 77(2025), 2 vom: 20. Feb., Seite 56-65
Auteur principal: Kinami, Ryuhei (Auteur)
Autres auteurs: Ineno, Toshinao
Format: Article en ligne
Langue:English
Publié: 2025
Accès à la collection:BioTechniques
Sujets:Journal Article Alkaline dip DNA extraction fish skin mucus genetic sexing high-throughput genotyping nucleic acid amplification point-of-care testing sturgeon swab sampling DNA plus... 9007-49-2 Sodium Hydroxide 55X04QC32I
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Résumé:Studies on simple DNA extraction methods from fish mucus, which is an ideal resource for noninvasive sampling, are scarce. In the aquaculture of sturgeons such as sterlets (Acipenser ruthenus), a high-throughput genetic sexing method is needed, as only females are reared to maturity for their roe. Here, DNA extraction methods using HotSHOT (hot sodium hydroxide and tris), a novel alkaline dip, alkaline glycol, and water were compared using the skin mucus of sterlets (n = 8) collected with a toothpick. High-throughput sexing of sterlets from six production batches (n = 3953) was also evaluated using HotSHOT or alkaline dip. An alkaline dip with 10-25 mM NaOH was an effective alternative to HotSHOT, eliminating the heating and neutralizing steps. Regarding high-throughput sexing, > 99% (3946) individuals were successfully genotyped on the first PCR trial, and genotype ratios of each batch were close to 1:1, showing applicability of the alkaline dip method for practical sterlet sexing
Description:Date Completed 09.04.2025
Date Revised 09.04.2025
published: Print-Electronic
Citation Status MEDLINE
ISSN:1940-9818
DOI:10.1080/07366205.2025.2467584