In a genetic improvement programme for strawberry (Fragaria × ananassa) it is important to have the methodology to assess the genetic integrity of the plant at all increment stages, from different criteria such as physiological, morphological and molecular; for this purpose, one of the most appropriate tools is the molecular markers called Simple Simple Repeats (SSR) Repeats Simple Simples, as they allow, for example, the identification of populations with reduced genetic diversity, the revealing of genealoogies, the knowledge of the degree of relationship between individuals, the provision of solid elements in the defence of intellectual property, the assessment of the purity of the plant material, the identification of the degree of somaclonal variation and the avoidance of mixing of plant material in germoplasma banks. In this regard, the aim of this study was to obtain the genetic footprint of strawberry varieties CP0201, CP0204, CP0615, CPLE7 developed in the College of Postgraduates and the Festival variety, developed in Florida, USA, with the use of nine microsatellite loci (SSR). The process included DNA extraction from strawberry foliar tissue as well as PCR end-point amplification of SSR loci grouped in multiplex reactions. PCR products were separated by capillary electrophoresis and their size in base pairs was determined by GeneMapper ® v. 4. From the allelic frequencies, the distance matrices were calculated using the coefficients of Jaccard and Dice. 63 different alleles were found, each pair of initiators amplified between 3 and 12 alleles. Markers with the highest number of alleles were EMFn181 (11 alleles) and EMFv104 (12 alleles). The genetic footprint of each variety was generated. The allelic profiles found differences between the varieties CP0615, CPLE7 and Festival; CP0201 and CP0204 had a similar genetic footprint as they are related through their female parent; the allelic diversity index within populations varied from 3.96 to 5.93. The varieties had a low rate of uniformity due to the high level of polymorphism of the markers used.