The purpose of the present study was to develop an effective and dependable protocol for in vitro regeneration of potato genotypes Lady Rosetta, Hermes, Cara, and Spunta. For the regeneration of potato genotypes, young shoots were collected, sterilized with varying concentrations of Clorox and 0.1% (w/v) HgC1₂, and then cultured on MS medium. For germplasm preservation and synthetic seed production, in vitro-grown plants were harvested. All genotypes were sterilized with 20% Clorox and 0.1% HgCl₂, which resulted in high survival rates. Several parameters, such as the culture medium, were modified to reduce growth. For long-period conservation, potato node cuttings stored in MS medium fortified with ABA 0.50 and 0.75 mg/l at 4 °C for 12 months without subculture gave 100% regrowth in MS medium containing 1 mg/l KN and normal growth conditions. This study showed the effect of storage potato shoots in a tissue culture medium containing ABA on DNA instability of potato genotypes, using the molecular markers RAPD and SSR. All RAPD and SSR primers showed differences in DNA bands. The use of single-stem cutting for synthetic seeds provides maintenance and transport advantages. In a 5% alginate gel, stem cuttings were encapsulated. As the concentration of ABA increased from 0.25 mg/l to 0.75 mg/l, the percentage of synthetic seeds that germinated and were converted into plantlets decreased gradually. The protocol can be used for exchanging potato germplasm and preparing synthetic seeds.