Skip to main content
SpringerLink
Log in

A simple and reliable method for DNA extraction from bivalve mantle

  • Short Communication
  • Published:
Journal of Applied Genetics Aims and scope Submit manuscript

Abstract

A simple and reliable method was developed for extracting genomic DNA from preserved mantle tissues of Pacific oysterCrassostrea 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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  • Aranishi F, 2006. A novel mitochondrial intergenic spacer reflecting population structure of Pacific oyster. J Appl Genet 47: 119–123.

    Article  PubMed  Google Scholar 

  • Aranishi F, Okimoto T, 2004. Genetic relationship between cultured populations of Pacific oyster revealed by RAPD analysis. J Appl Genet 45: 435–443.

    PubMed  Google Scholar 

  • Aranishi F, Okimoto T, 2005. Sequence polymorphism in a novel noncoding region of Pacific oyster mitochondrial DNA. J Appl Genet 46: 201–206.

    PubMed  Google Scholar 

  • Berthelin C, Kellner K, Mathieu M, 2000. Storage metabolism in the Pacific oyster (Crassostrea gigas) in relation to summer mortalities and reproductive cycle (west coast of France). Comp Biochem Physiol 125B: 359–369.

    CAS  Google Scholar 

  • Brokordt KB, Himmelman JH, Guderley HE, 2000. Effect of reproduction on escape responses and muscle metabolic capacities in the scallopChlamys islandica Muller 1776. J Exp Mar Biol Ecol 251: 205–225.

    Article  PubMed  Google Scholar 

  • Encomio V, Chu FL, 2000. The effect of PCBs on glycogen reserves in the eastern oysterCrassostrea virginica. Mar Environ Res 50: 45–49.

    Article  CAS  PubMed  Google Scholar 

  • Fernandez A, Garcia T, Asensio L, Rodriguez MA, Gonzalez I, Cespedes A, et al. 2000. Identification of the clam speciesRuditapes decussatus (grooved carpet shell),Venerupis pullastra (pullet carpet shell), andRuditapes philippinarum (Japanese carpet shell) by PCR-RFLP. J Agric Food Chem 48: 3336–3341.

    Article  CAS  PubMed  Google Scholar 

  • Hyotylainen T, Karels A, Oikari A, 2002. Assessment of bioavailability and effects of chemicals due to remediation actions with caging mussels (Anodonta anatina) at a creosote-contaminated lake sediment site. Water Res 36: 4497–4504.

    Article  CAS  PubMed  Google Scholar 

  • Klinbunga S, Ampayup P, Tassanakajon A, Jarayabhand P, Yoosukh W, 2001. Genetic diversity and molecular markers of cupped oysters (generaCrassostrea, Saccostrea, andStriostrea) in Thailand revealed by randomly amplified polymorphic DNA analysis. Mar Biotechnol 3: 133–144.

    Article  CAS  PubMed  Google Scholar 

  • Ko YT, Chan MMY, Ford SE, Fong D, 1999. A PCR-ELISA method for direct detection of the oyster pathogenHaplosporidium nelsoni. Mar Biotechnol 1: 147–154.

    Article  CAS  PubMed  Google Scholar 

  • Launey S, Hedgecock D, 2001. High genetic load in the Pacific oysterCrassostrea gigas. Genetics 159: 255–265.

    CAS  PubMed  Google Scholar 

  • Martinez G, Brokordt K, Aguilera C, Soto V, Guderley H, 2000. Effect of diet and temperature upon muscle metabolic capacities and biochemical composition of gonad and muscle inArgopecten purpuratus Lamarck 1819. J Exp Mar Biol Ecol 247: 29–49.

    Article  CAS  PubMed  Google Scholar 

  • Neudecker F, Grimm S, 2000. High-throughput method for isolating plasmid DNA with reduced lipopolysaccharide content. Biotechniques 28: 106–108.

    Google Scholar 

  • Sato M, Nagashima K, 2001. Molecular characterization of a mitochondrial DNA segment from the Japanese scallop (Patinopecten yessoensis): Demonstration of a region showing sequence polymorphism in the population. Mar Biotechnol 3: 370–379.

    Article  CAS  PubMed  Google Scholar 

  • Steiner G, Muller M, 1996. What can 18S rDNA do for bivalve phylogeny? J Mol Evol 43: 58–70.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological and Environmental Sciences, Miyazaki University, 889-2192, Miyazaki, Japan

    Futoshi Aranishi & Takane Okimoto

Authors
  1. Futoshi Aranishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takane Okimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Futoshi Aranishi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aranishi, F., Okimoto, T. A simple and reliable method for DNA extraction from bivalve mantle. J Appl Genet 47, 251–254 (2006). https://doi.org/10.1007/BF03194632

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03194632

Key words

Search

Navigation