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Efficient characterization of biological diversity using field DNA extraction and random amplified polymorphic DNA markers

D.J. Fairbanks^I; A. Waldrigues^II; C.F. Ruas^II; P.M. Ruas^II; P.J. Maughan^III; L.R. Robison^III; W.R. Andersen^I; C.R. Riede^IV; C.S. Pauley^I; L.G. Caetano^II; O.M.N. Arantes^II; M.H.P. Fungaro^II; M.C. Vidotto^V; S.E. Jankevicius^V

^IDepartment of Botany and Range Science, Brigham Young University, Provo, Utah 84602, USA. Send correspondence to D.J.F.
^IIDepartamento de Biologia Geral, Universidade Estadual de Londrina, Caixa Postal 6001, 86051 Londrina, PR, Brasil
^IIIDepartment of Agronomy and Horticulture, Briham Young University, Provo, Utah 84602, USA
^IVInstituto Agronômico do Paraná, Caixa Postal 1331, 86001 Londrina, PR, Brasil
^VDepartamento de Microbiologia, Universidade Estadual de Londrina, Caixa Postal 6001, 86051 Londrina, PR, Brasil

ABSTRACT

DNA polymorphism analyses, such as restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analyses, are particularly useful for genetic characterization of biological resources. We describe procedures for DNA extraction and RAPD analysis that are adapted for analyzing large numbers of samples with minimal laboratory equipment. The DNA extraction procedure reported here provided ample amounts of DNA of the purity required for RFLP or RAPD analyses from various plant, animal, and protozoan species. The procedure requires relatively small amounts of tissue, does not require phenol or chloroform, and may be conducted under field conditions in the absence of running water and electricity. RAPD markers are particularly valuable when compared to RFLPs or isozymes since decanucleotide primers of arbitrary sequence tend to produce a relatively high frequency of polymorphisms per primer and the procedure is rapid and cost efficient. Reduction of amplification reaction volume from 25 to 15 mI resulted in amplified DNA markers at a 33% reduction in cost. Using quinoa (Chenopodium quinoa) genetic resources as a model, a total of 95 primers were initially screened on a single genotype to select those primers that amplified

Keywords: Biological diversity; Polymorphic DNA markers.

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