Conditions for amplification microsatellites in potato cultivars

Abstract

The identification of cultivars of potato, Solanum tuberosum L., is critical for the management of genebanks, breeding programs, genetic diversity analysis, processes micropro­ pagation and seed production. Because of its high degree of polymorphism, microsatellites (SSR) have proved a powerful tool for analysis of variability is also reproducible. The objec­ tive was to standardize the conditions for DNA amplification using four accessions from the germplasm bank of INIA- Merida using microsatellite markers. DNA extraction was performed with a fast, economical and reliable method. Of the 24 primers proposed diversity studies were selected for their higher content polimórfico12 markers. The amplified products were observed in gels of 3% agarose, stained with ethidium bromide. We tested different methods used in previous studies for genetic diversity and identity in potato, is it possible to obtain DNA fragment patterns sharp, accurate and reproducible one whose optimum concentrations was 0.2 U of Taq polymerase, 1.5 mM MgCl2 and 0.2 µM of primers and 30 min decreased over time in each amplification,  yielding greater efficiency per reaction. Of the 12 primers, four were selected based on their polymorphism and the quality of the amplified DNA fragments