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RAPD-PCR amplification of soybean DNA using pairwise combinations of primers
Ronan Xavier CorrêaI,III; Everaldo Gonçalves de BarrosI,III; Fábio Gelape FaleiroI,III; Maurilio Alves MoreiraII,III
IDepartamento de Biologia Geral, Universidade Federal de Viçosa, 36571-000 Viçosa, MG, Brasil.
IIDepartamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, 36571-000 Viçosa, MG, Brasil.
IIINúcleo de Biotecnologia Aplicada à Agropecuária (BIOAGRO), Universidade Federal de Viçosa, 36571-000 Viçosa, MG, Brasil. Send correspondence to E.G.B.
ABSTRACT
In order to increase the number of detectable polymorphisms for the construction of a RAPD-PCR based intraspecific soybean genetic map, two primers were used at a time, instead of only one. The DNA patterns obtained with each combination of primers were analyzed for intensity and number of bands, as well as for reproducibility. DNA samples of two soybean progenitors were tested initially with 72 random primers and 36 random pairwise combinations of these primers. Polymorphic fragments generated with the combinations of primers segregated 3 to 1 in an F2 population of 68 individuals derived from these progenitors. Linkage analysis of some of the polymorphic bands revealed that they did not co-segregate with bands generated with the isolated primers. Our data show that pairwise combinations of primers can facilitate the detection of RAPDs.
Keywords: RAPD-PCR; soybean DNA.
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