Abstract:By selecting 22 pairs of polymorphic microsatellite markers of Portunus trituberculatus, primers matching, and combinatorial testing, the multiplex PCR amplification system was built and the reaction conditions were optimized, including annealing temperature, Mg2+ concentration, dNTPs concentration and other parameters. Forty-seven combinations of double PCR, 15 combinations of triple PCR, and 3 quadruple PCR combinations were established. The establishment of this technology will provide a fast, effective molecular detecting tool for the P. trituberculatus breeding and germ-line assessment.
