Abstract
In order to avoid the effects of bacterial contamination and the excess of RNA and polysaccharides coming from the cell walls, algal DNA for PCR cycling in RAPD analysis was extracted from protoplasts and purified in a CsCl gradient. Results indicate that RAPD can be efficiently applied to marine algæ and useful to distinguish between different strains or between different species.
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References
Butler, D.M., L.V. Evans, and B. Kloareg. 1990. Isolation of protoplasts from marine macroalgæ. In: Akatska I. (Ed.),Introduction to Applied Phycology. SPB Acad-Publ. The Hague, The Netherlands, pp. 647–668.
Dellaporta, S.L., J. Wood and J.B. Hicks 1983. A plant DNA minipreparation version II. Plant Mol. Biol. Reporter 1:19–21.
Loiseaux, S., and C. Rozier. 1978. Culture axénique dePylaiella littoralis (L.) Kjellm. (Phéophycées). Rev. Algol. N.S. 13:333–340.
Mejjad, M., S. Loiseaux de Goer, and G. Ducreaux. 1992. Protoplast isolation, development, and regeneration in different strains ofPylaiella littoralis (L.) Kjellm. (Phæophyceæ). Protoplasma 169:42–48.
Müller, D.G. and B. Stache. 1989. Life history studies onPilayella littoralis (L.) Kjellman (Phæophyceæ, Ectocarpales) of different geographical origin. Bot. Mar. 32:71–78.
Murray, H.G. and W.F. Thompson. 1980. Rapid isolation of high molecular weight DNA. Nucleic Acids Res. 8:4321–4325.
Patwary, M.U., R.M. Mackay and J.P. van der Meer. 1993. Revealing genetic markers inGelidium vagum (Rhodophyta) through the random amplified polymorphic DNA (RAPD) technique. J. Phycol. 29:216–222.
Pikaart, M.J., and B. Villeponteau. 1993. Suppression of PCR amplification by high levels of RNA. Biotechniques 14:24–25.
Richards, E.J., and F.M. Ausubel. 1988. Isolation of a higher eukaryotic telomere fromArabidopsis thaliana. Cell 53:127–136.
Saghai-Maroof, M.A., K.M. Soliman, R.A. Jorgensen, and R.W. Allard. 1984. Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location and population dynamics. Proc. Natl. Acad. Sci. USA 81:8014–8018.
Saiki, R.K., D.H. Gelfand, S. Stoffel, S.J. Sharf, R. Higachi, G.T. Horn, K.B. Malles and H.A. Elrich. 1988. Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491.
San, L.H., F. Vedel, D. Sihachakr and R. Rémy. 1990. Morphological and molecular characterization of fertile tetraploïd somatic hybrids produced by protoplast electrofusion and PEG-induced fusion betweenLycopersicon esculentum Mill. andLycopersicon peruvianum Mill. Mol. Gen. Genet. 221:17–26.
Williams, J.G.K., A.R. Kabelik, K.J. Livak, J.A. Rafalski and S.V. Tingey. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Res. 18:6531–6535.
Yoon, C.S., and D.A. Glawe. 1993. Pretreatment with RNase to improve PCR amplification of DNA using 10-mer primers. Biotechniques 14:908–910.
Zobell, C.E. 1941. Studies on marine bacteria. I. The cultural requirements of heterotrophic aerobes. J. Mar. Res. 4:41–75.
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A detailed version of the protocols employed is published in the electronic edition of theReporter: PMBR.ElectEd.12(2).
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Mejjad, M., Vedel, F. & Ducreux, G. Improvement of DNA preparation and of PCR cycling in RAPD analysis of marine macroalgæ. Plant Mol Biol Rep 12, 101–105 (1994). https://doi.org/10.1007/BF02668370
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DOI: https://doi.org/10.1007/BF02668370