Development of a Multiplex Microsatellite PCR Assay Based on Microsatellite Markers for the Mud Carp,Cirrhinus molitorella

In this study, we developed a multiplex microsatellite polymerase chain reaction (PCR) assay that consisted of 13 pairs of primers selected from previously developed microsatellite markers in mud carp data obtained in our laboratory. We adjusted the number of cycles, the time of extension, and the primer ratios to make the multiplex PCR system as valuable as possible. Using this system, we performed a genetic analysis and parentage assignment for 2 male and 5 female mud carps and 146 of their progeny. The results showed a range of alleles of 2 < K < 5, with an average of 3.54. The polymorphism information content ranged from 0.271 to 0.697, with an average of 0.4954. The observed heterozygosity ranged from 0.183 to 0.810, with an average of 0.6008. The expected heterozygosity ranged from 0.324 to 0.741, with an average of 0.5572. The null allele frequency was 1.29–27.68%. The computer‐simulated identification was 99%, while the combined exclusion power using all loci was 100% for actual offspring. Thus, this multiplex PCR assay is a new technological tool for selective breeding and a genetic resource for studying the mud carp, Cirrhinus molitorella.