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Brassica oleracea var. capitata L. is characterized by a high level of intraspecific heterogeneity due to some biological features that cause difficulties for breeding creating genetically homogenous forms and maintaining their genetic purity. Microsatellites (SSRs) are highly polymorphic markers of plant genomes and represent one of the most effective tools for assessing genetic polymorphism. Among microsatellites, EST-SSRs are the most interesting, because they are directly linked to the expressed sequences and for that reason are widely used for the analysis of genetic diversity and population structure. In this work, we studied the effectiveness of the use of transferable EST-SSR markers for analyzing both white cabbage diversity and genotyping of pure lines. As a result, 15 microsatellite loci were characterized for their information content, allelic frequencies, and heterogeneity levels. The effective multiallelic markers (Bo20TR, BoDCTD4, BoPC34, BoPLD1, BoCalc and BoPC15) with high information content (PIC > 0.7) that could be successfully used for the analysis of inter- and intravarietal polymorphism in Brassica oleracea var. capitata were identified. It has been shown that intervarietal polymorphism expressed as the allelic diversity of the EST-SSR loci greatly facilitates varietal identification and typing of individual plants for breeding purposes. Based on the SSR-evaluation and subsequent clustering, the genetic structure of the breeding collection was identified, which showed that most experimental forms, in spite of their different origins, have a common ancestral genetic basis. The identified donors of rare alleles could potentially be a source of valuable genetic segregation for further B. oleracea breeding improvement.