**5. Conclusions**

In summary, two multiplex PCR amplification systems were constructed using 12 microsatellite markers to establish a quasi-rapid and efficient microsatellite analysis technique for *C. undulatus*. The constructed multiplex amplification systems were stable, and the loci were highly polymorphic, providing a basis for germplasm identification, genetic diversity analysis, and stocking effect evaluation of *C. undulatus*. This method can be used to select a set of primers with high polymorphism and stable PCR amplification from the developed microsatellite primers and establish an accurate, rapid, and efficient microsatellite analysis technique using multiplex PCR combination, which can provide technical support for population genetics and family lineage analysis of species.

**Author Contributions:** Conceptualization, D.Z.; methodology, F.Z. and L.G.; software, L.G.; validation, F.Z. and J.Y.; formal analysis, F.Z.; resources, N.Z. and K.Z.; data curation, F.Z.; writing—original draft preparation, F.Z.; visualization, F.Z.; supervision, H.G. and K.Z.; project administration, L.G. and B.L. All authors have read and agreed to the published version of the manuscript.

**Funding:** This study was funded by Central Public-Interest Scientific Institution Basal Research Fund, CAFS (2020GH06), and Financial Fund of the Ministry of Agriculture and Rural Affairs, P. R. of China (NHYYSWZZZYKZX2020), and National Marine Genetic Resource Center, and China-ASEAN Maritime Cooperation Fund.

**Institutional Review Board Statement:** All applicable international, national, and institutional guidelines for the care were followed by the authors.

**Informed Consent Statement:** Informed consent was obtained from all subjects involved in the study. Written informed consent has been obtained from the patients to publish this paper.

**Data Availability Statement:** All data generated or analyzed during this study are included in this published article.

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**

