Abstract
Laver (Porphyra yezoensis) DNAs were extracted from thalli with five different procedures and used for RAPD (random amplified polymorphic DNA) analysis as templates. Restriction enzyme-digestive DNAs were obtained with all procedures examined. However, RAPD patterns generated with these DNAs appeared highly irreproducible and were considerably different from each other. When DNAs purified with CsCl gradient centrifugation were used for RAPD analysis as templates, highly reproducible RAPD patterns were obtained, suggesting that unpurified DNAs extracted from thalli with all five extraction procedures contained an excess of RNA, polysaccharides and/or other materials which affected the RAPD reproducibility. Thus, results indicated that purification of DNA is essential to produce reproducible RAPD patterns of Porphyra DNA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abo K, Noda H (1995) Isozymes from conchocelis of Porphyra species. Fisheries Sci. 61: 249–251.
Fujita Y, Migita S (1987) Fusion of protoplasts from thalli of two different color types in Porphyra yezoensis U. and development of fusion products. Jpn. J. Phycol. 35: 201–208.
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Micheli MR, Bova R, Pascal E, D'Ambrosio E (1994) Reproducible DNA fingerprinting with the random amplified polymorphic DNA (RAPD) method. Nucleic Acids Res. 22: 1921–1922.
Miura A, Fu PF, Shin JA (1992) Interspecific cross experiments between Porphyra yezoensis Ueda and P. tenera Kjellman (Bangiales, Rhodophyta) by using pigmentation variants. J. Tokyo Univ. Fish. 79: 103–120 (in Japanese with English abstract).
Mizukami Y, Okauchi M, Kito H, Ishimoto S, Ishida T, Fuseya M (1995) Culture and development of electrically fused protoplasts from red marine algae, Porphyra yezoensis and P. suborbiculata. Aquaculture 132: 361–367.
Mizukami Y, Okauchi M, Kito H, Kobayashi M (1996a) DNA fingerprinting of cultivated laver (Porphyra yezoensis and P. tenera) with oligonucleotide probes, and its application on cultivar discrimination. Fisheries Sci. 62: 173–177.
Mizukami Y, Okauchi M, Kito H, Kobayashi M (1996b) Discrimination of laver cultivars with RAPD markers. Fisheries Sci. 62: 547–551.
Pandey RN, Adams RP, Flournoy LE (1996) Inhibition of random amplified polymorphic NAs (RAPDs) by plant polysaccharides. Plant Mol. Biol. Reporter 14: 17–22.
Patway MU, van der Meer JP (1994) Application of RAPD markers in an examination of heterosis in Gelidium vagum (Rhodophyta). J. Phycol. 30: 91–97.
Pikaart MJ, Villeponteau B (1993) Suppression of PCR amplification by high levels of RNA. Biotechniques 14: 14–25.
Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with abritrary primers. Nucleic Acids Res. 18: 7213–7218.
Williams JGK, Kubelik AE, Levak KJ, Rafalski JA, Tingey SC (1990) DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531–6535.
Zhao X, Kochert G (1992) Characterization and genetic mapping of a short highly repeated interspersed DNA sequence from rice (Oryza sativa L.). Mol. Gen. Genet. 231: 353–359.
Yoon CS, Glave DA (1993) Pretreatment with RNase to improve PCR amplification of DNA using 10-mer primers. Biotechniques 14: 908–910.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mizukami, Y., Kito, H., Kunimoto, M. et al. Effect of DNA preparation from laver (Porphyra yezoensis) thalli on reproducibility of RAPD (random amplified polymorphic DNA) patterns. Journal of Applied Phycology 10, 23–29 (1998). https://doi.org/10.1023/A:1008023806500
Issue Date:
DOI: https://doi.org/10.1023/A:1008023806500