Shake and stew: a non-destructive PCR-ready DNA isolation method from a single preserved fish larva

Shake and stew: a non-destructive PCR-ready DNA isolation method from a single preserved fish larva

A rapid non‐destructive alternative to isolate DNA from an individual fish larva is presented, based on the suspension of epithelial cells through vortex forces, and the release of DNA in a heated alkaline solution. DNA from >6056 fish larvae isolated using this protocol has yielded a high PCR am...

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Veröffentlicht in:Journal of fish biology 2014-01, Vol.84 (1), p.267-272
Hauptverfasser: Alvarado Bremer, J. R., Smith, B. L., Moulton, D. L., Lu, C.-P., Cornic, M.
Format: Artikel
Sprache:eng
Schlagworte:
Animals
Deoxyribonucleic acid
DNA
DNA - isolation & purification
Epithelial Cells
Fishes
genetics
ichthyoplankton
Istiophorus platypterus
Larva - genetics
Microbiology
minimally invasive
NaOH
Polymerase Chain Reaction - methods
Specimen Handling - methods
Thunnus albacares
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Beschreibung
Zusammenfassung:A rapid non‐destructive alternative to isolate DNA from an individual fish larva is presented, based on the suspension of epithelial cells through vortex forces, and the release of DNA in a heated alkaline solution. DNA from >6056 fish larvae isolated using this protocol has yielded a high PCR amplification success rate (>93%), suggesting its applicability to other taxonomic groups or sources when tissue amount is the limiting factor.
ISSN:0022-1112
1095-8649
DOI:10.1111/jfb.12280