Abstract
For genetic analysis of fungal DNAs, we have modified the RAPD method to use primers with G+C contents of 80–100%. In RAPD analysis of Puccinia graminis f. sp. tritici DNAs, these primers generated twice the number of both amplification products per primer and polymorphisms among isolates as compared to the standard 60–70% G+C primers. With respect to segregation and genetic similarity, RAPD markers generated by the high-GC primers behaved as do RAPD markers produced by the standard primers. These high-GC primers also yielded increased numbers of amplification products in RAPDs on the DNAs of a broad range of other fungi.
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References
Backlund JE, Szabo LJ (1993) Physical characteristics of the genome of the phytopathogenic fungus Puccinia graminis. Curr Genet 24:89–93
Boehm EWA, Wenstrom JC, McLaughlin DJ, Szabo LJ, Roelfs AP, Bushnell WR (1992) An ultrastructural pachytene karyotype for Puccinia graminis f.sp. tritici. Can J Bot 70:401–413
Crowhurst RN, Hawthorne BT, Rikkerink EHA, Templeton MD (1991) Differentiation of Fusarium solani f.sp. cucurbitae races 1 and 2 by random amplification of polymorphic DNA. Curr Genet 20:391–396
Goodwin PH, Annis SL (1991) Rapid identification of genetic variation and pathotype of Leptosphaeria maculans by random amplified polymorphic DNA assay. Appl Environ Microbiol 57:2482–2486
Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MAPMAKER: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181
Liu Z, Szabo LJ, Bushnell WR (1993) Molecular cloning and analysis of abundant and stage-specific mRNAs from Puccinia graminis. Mol Plant-Microbe Interact 6:84–91
Roelfs AP (1985) Wheat and rye stem rust. In: Bushnell WR, Roelfs AP (eds). The Cereal Rusts, vol II. Academic Press, Orlando, pp 3–37
Rohlf FJ (1993) NTSYS-pc. Numerical taxonomy and multivariate analysis system. Exeter Software, Setauket, New York
Weeden NF, Timmerman GM, Hemmat M, Kneen BE, Lodhi MA (1992) Inheritance and reliability of RAPD markers. In: Applications of RAPD Technology to Plant Breeding. Proceedings of the Symposium on Joint Plant Breeding, pp 12–17
Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
Williams JGK, Hanafey MK, Rafalski JA, Tingey SV (1993) Genetic analysis using random amplified polymorphic DNA markers. Methods Enzymol 218:704–740
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Communicated by O.C. Yoder
Present address: Department of Biology, Hiram College, Hiram, OH 44234, USA
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Kubelik, A.R., Szabo, L.J. High-GC primers are useful in RAPD analysis of fungi. Curr Genet 28, 384–389 (1995). https://doi.org/10.1007/BF00326438
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DOI: https://doi.org/10.1007/BF00326438