Hydrobiologia

, Volume 359, Issue 1–3, pp 159–162 | Cite as

Extraction of DNA from anostracan cysts (Crustacea, Branchiopoda) for use in RAPD-PCR analyses

  • Jacob A. Moorad
  • Michael S. Mayer
  • Marie A. Simovich

Abstract

Like many diapausing crustaceans, anostracans (Crustacea, Branchiopoda) produce encysted embryos capable of surviving the long periods of desiccation typical of their environments. These cysts are far more abundant and are easier to collect than adults and for some applications they are superior to adults as subjects for genetic analysis. Due to the lack of minimal tissue size requirements, PCR-based analysis is the best alternative for genetic study of cysts. We describe a method for the fast extraction of DNA from cysts, yielding template for RAPD-PCR analysis.

Crustacea Anostraca cysts egg bank RAPD PCR 

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References

  1. Belk, D., 1977. Evolution of egg size strategies in fairy shrimps. S. West Nat. 22: 99–105.Google Scholar
  2. Clegg, J. S. & F. P. Conte, 1980. A review of the cellular and developmental biology of Artemia. In Persoone, G., P. Sorgloos, O. Roels & E. Jaspers (eds). The Brine Shrimp Artemia. Vol. 2. Physiology, Biochemistry, Molecular Biology. Universa Press, Wetteren, Belgium: 11–54.Google Scholar
  3. Davis, L. G., M. D. Dibner & J. F. Battey, 1986. Basic Methods in Molecular Biology. Elsevier Science Publishing Co., Inc., New York, 388 pp.Google Scholar
  4. Lécher, P., D. Defaye & P. Noel, 1995. Chromosomes and nuclear DNA of Crustacea. Invert. Reprod. Dev. 27: 85–114.Google Scholar
  5. Linder, H. J., 1960. Studies on the fresh water fairy shrimp Chirocephalopsis bundyi(Forbes). II. Histochemically of EggShell Formation. J. Morph. 107: 259–284.PubMedCrossRefGoogle Scholar
  6. Moorad, J.A, M.A. Simovich & M. S. Mayer, 1997. Identification of Southern Californian Branchinectid Cysts (Crustacea, Anostraca) using RAPD-PCR Species-Specific Markers. J. Crust. Biol. (In Press).Google Scholar
  7. Saiki, R., D. H. Gelfand, S. Stoffel, S. J. Scharf, R. Higuchi, G. T. Horn, K. B. Mullis & H. A. Erlich, 1988. Primerdirected enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487–491.PubMedGoogle Scholar
  8. Simovich, M. A. & S. A. Hathaway, 1997. Diversified bet-hedging as a reproductive strategy of some ephemeral pool anostracans (Branchiopoda). J. Crust. Biol. 17: 38–44.CrossRefGoogle Scholar
  9. Walsh, P. S., D. A. Metzger & R. Higuchi, 1991. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. BioTechniques 10: 506–513.PubMedGoogle Scholar
  10. Welsh, J. & M. McClelland, 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 18: 7213–7218.PubMedGoogle Scholar
  11. Williams, J. G. K., A. R. Kubelik, K. J. Livak, J. A. Rafalski & S. V. Tingey, 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531–6535.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Jacob A. Moorad
    • 1
  • Michael S. Mayer
    • 2
  • Marie A. Simovich
    • 2
  1. 1.Department of Marine and Environmental ScienceUniversity of San DiegoUSA
  2. 2.Department of BiologyUniversity of San DiegoUSA

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