Detection of minisatellite sequences inPhaseolus vulgaris |
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Authors: | T Stockton G Sonnante P Gepts |
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Institution: | (1) Department of Agronomy and Range Science, University of California, 95616-8515 Davis, CA, USA |
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Abstract: | An improved procedure for detecting minisatellite sequences inPhaseolus vulgaris is described. Both M13 protein III tandem repeat and the 33.15 human mini-satellite sequences revealed polymorphisms with
a high number of sharp bands after digestion of genomic DNA withHae III,Hinf I, orTaq I. Improved resolution of the numerous restriction fragments detected by these probes is accomplished by one or more of the
following: varying agarose concentration, using high SDS hybridization buffer, exposure of the autoradiograph without intensifying
screens, and transfer of the autoradiograph to electrophoresis paper. Increased stability of the DNA-DNA hybridizations with
these heterologous probes is obtained by reducing hybridization temperature. Labeling probes with the polymerase chain reaction
can accentuate some restriction fragments depending upon the radiolabeled nucleotide used. |
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Keywords: | Common bean RFL M13 fingerprint |
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