Electrophoretic karyotyping without the need for generating protoplasts

Abstract

Chromosome samples for pulsed-field electrophoresis have been prepared without first generating protoplasts. The technique involves treatment of intact, agarose-solidified cell material with protease in the presence of EDTA and SDS. Saccharomyces cerevisiae, Ustilago hordei, Tilletia caries, and T. controversa karyotypes are clearly resolved with this technique. Colonies of U. hordei and S. cerevisiae removed from the surface of agarsolidified media and prepared for PFGE by this abbreviated method also yield well resolved karyotypes. © 1990 Springer-Verlag.