A simple and reliable method for DNA extraction from bivalve mantle.
A simple and reliable method was developed for extracting genomic DNA from preserved mantle tissues of Pacific oyster Crassostrea gigas for reproducible PCR amplification. The method was based on destruction of the tissue using Proteinase K, Chelex 100 resin, detergents, and urea, followed by preferential capturing of genomic DNA with silica particles. Approximately 5 mg of mantle tissue provided a sufficient quality and quantity of DNA for several hundreds of PCR reactions amplifying the hypervariable mitochondrial DNA intergenic spacer, which is a useful genetic marker for population structure analysis of Pacific oyster. The method can be applied for DNA preparation from not only fresh and frozen but also ethanol-preserved mantle tissues, so this rapid and economical method can serve for investigating a large number of bivalve specimens collected in the field and next transported in ethanol at ambient temperature.
Aranishi
Futoshi
F
Department of Biological and Environmental Sciences, Miyazaki University, Miyazaki 889-2192, Japan. aranishi@cc.miyazaki-u.ac.jp
Okimoto
Takane
T
D000818:Animals; D049896:Crassostrea / Q000235:genetics*; D004247:DNA / Q000302:isolation & purification*; D005821:Genetic Techniques; D016133:Polymerase Chain Reaction
D004247:DNA