A microsatellite marker parentage identification primer, method and application suitable for Nile tilapia, Olia tilapia and their hybrids

[0031] A kind of microsatellite marker parentage identification method applicable to Nile tilapia, Olia tilapia and hybrids thereof, comprising the steps:

[0032] 1. Family construction: Using the parents of Nile tilapia and Olia tilapia preserved in this laboratory, after they are sexually mature, use artificial insemination to construct 3 Nile tilapia full-sib families, 2 A full-sib family of Olia tilapia and three full-sib families of Olia tilapia (Nile tilapia♀×Olia tilapia♂). Each family was numbered and reared independently, and 2 months after the fry hatched, the tail fin samples of the parents of each family and 32 to 35 offspring were collected as materials for paternity identification.

[0033] 2. Genomic DNA extraction: The caudal fin genomic DNA of the parent and offspring tilapias of each family in step 1 was extracted using an animal tissue micro DNA extraction kit (Magen).

[0034] 3. Screening of polymorphic microsatellite primers: 2 to 3 microsatellite loci ...

[0052] Practical application effect evaluation: paternity identification of tilapia "Yuemin No. 1" family

[0053] 1. Family construction and sampling: 5 female Nile tilapias were selected as female parents, and 5 super-male Nile tilapias were selected as male parents and Olia tilapia as female parents. This, the hybrid offspring obtained after backcrossing with the super-male Nile tilapia) was used as the male parent, and 5 full-sib families were constructed by one-to-one pairing, and the parents and 35 offspring of each family were collected after natural seedling production tail fin samples.

[0054] 2. Using the method of step 2 in Example 1 to extract the genomic DNA of the parents and offspring of the full-sib family of the above-mentioned 5 tilapia "Yuemin No. 1"; adopting the method of step 4 in Example 1 to adopt 13 pairs of fluorescently labeled microsatellites The primers were used for PCR amplification and ABI 3730xl sequencer for multiple capillary electrophoresi...