Optimization of the Analysis of Almond DNA Simple Sequence Repeats (SSRs) Through Submarine Electrophoresis Using Different Agaroses and Staining Protocols

摘要: Simple sequence repeat (SSR markers or microsatellites), based on the specific PCR amplification of DNA sequences, are becoming choice for molecular characterization a wide range plants because their high polymorphism, abundance, and codominant inheritance. Different methods have been used analysis SSR amplified fragments being submarine agarose electrophoresis more suitable method routine application. In this work we performed comparative study utilization four different types low melting (Metaphor®, Sea Kem®, MS-8®) regular (LD-2®) agaroses two staining protocols using Ethidium Bromide Gel Red Nucleic Acid Sating®. Almond cultivars assayed included Spanish 'Antoneta', 'Marta', 'Penta', 'Tardona' 'Desmayo' 'Guara', French 'Ferragnes' 'R1000', USA cultivar 'Mission', Tunisian 'Achaak', Italian 'Tuono' Australian 'Chellaston'. detection Metaphor® gel was most efficient with higher resolution would be able to resolve allelic variation in comparison other three assayed. addition, showed similar results than GelRed TM Stain® although it is much toxic. The use MetaPhor® appears good indicated mapping population due its rest agaroses, less toxicity Bromide, lower cost easier application automatic capillary sequencing.