Plant Molecular Biology Reporter

, Volume 10, Issue 1, pp 47–59

Detection of minisatellite sequences inPhaseolus vulgaris

  • T. Stockton
  • G. Sonnante
  • P. Gepts
Protocol

DOI: 10.1007/BF02669264

Cite this article as:
Stockton, T., Sonnante, G. & Gepts, P. Plant Mol Biol Rep (1992) 10: 47. doi:10.1007/BF02669264

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.

Key Words

Common bean RFL M13 fingerprint 

Abbreviations

PCR

polymerase chain reaction

BSA

bovine serum albumin

SSC

150 mM NaCl, 15 mM Na citrate, pH 7.0

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • T. Stockton
    • 1
  • G. Sonnante
    • 1
  • P. Gepts
    • 1
  1. 1.Department of Agronomy and Range ScienceUniversity of CaliforniaDavisUSA

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