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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Veröffentlicht in:BioTechniques. - 1991. - 77(2025), 2 vom: 20. Feb., Seite 56-65
1. Verfasser: Kinami, Ryuhei (VerfasserIn)
Weitere Verfasser: Ineno, Toshinao
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2025
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte: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 mehr... 9007-49-2 Sodium Hydroxide 55X04QC32I
Beschreibung
Zusammenfassung: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
Beschreibung: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