Abstract:

AFLP(Amplified Fragment Length Polymorphism) analysis system for Oncorhyrchus masou masou were established in this study, the genomic DNA were digested by two groups of restriction enzymes, with the relative processes being optimized, including the quantity of the restricting enzymes, the time of digested reaction, quantity of the template for pre-amplification and times of pre-amplification products dilution. It was proved that the results were best with 100ng DNA digested for 3h at 37℃ by 3U EcoRI and then 4h at 65℃ by TaqI and Tru9I respectively, then ligated by ligase. Quite clearly results could be achieved by 3μL ligation products as the template for pre-amplification which products were diluted 50 times for selective amplification. The comparison of the two pairs showed that the group of EcoRI-Tru9I obtained more amplification bands but lower ratio of polymorphic loci than EcoRI-TaqI.