Single Fish Egg DNA Extraction for PCR Amplification

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書誌事項

公開日
2006-02
権利情報
  • http://www.springer.com/tdm
DOI
  • 10.1007/s10592-005-5387-y
公開者
Springer Science and Business Media LLC

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説明

Modern stock researches on marine biomass are basically genetic and rely increasingly on PCR-based manipulations of informative DNA markers for detecting the genetic diversity. This study developed a simple and rapid single tube method for DNA extraction from a single fish egg. The 15 min protocol was based on the use of Chelex 100 resin and urea to breakdown membrane and connective tissue of eggs. From various sizes of a single egg of walleye pollack (Theragra chalcogramma), the amounts of total nucleic acids were reproducibly obtained to be 18.25 ± 1.92 μg per egg. Using DNA templates diluted ranging 1/100–1/105, PCR amplification for the mitochondrial cytochrome b (Cytb) gene was successfully performed, and the 1/102 diluted template yielded the best result in PCR amplification for three different DNA marker genes. This method is quite simple and economical, and enables to provide the high throughput often demanded by the stock identification of marine biomass, in which large numbers of specimens of single fish eggs must be analyzed.

収録刊行物

  • Conservation Genetics

    Conservation Genetics 7 (1), 153-156, 2006-02

    Springer Science and Business Media LLC

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