Method utilizing microsatellite markers to identify polycultured Songpu mirror carp family
A microsatellite marker and polyculture technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of allelic differences, the inability of microsatellite markers to be universal and unsuitable for application, and achieve the effect of short cycle.
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specific Embodiment approach 1
[0015] Specific implementation mode one: this implementation mode identification polyculture Matsuura mirror carp family is carried out according to the following steps:
[0016] 1. Extract the individual genome DNA of the parents and offspring of each family of Matsuura mirror carp, and use the DNA of the parents and offspring as templates, and perform multiple PCR reactions with microsatellite marker primers;
[0017] 2. Load the PCR product of step 1 on a 10% polyacrylamide gel, perform polyacrylamide gel electrophoresis, and develop the color by silver staining. The electrophoretic pattern is analyzed by Gel-pro analyzer4.5 software, combining the length of the PCR product and the four bases The repeat unit confirms the size of the gene fragment and determines the genotype;
[0018] Three, set up the genotype data file of polyculture Matsuura mirror carp parent and progeny according to the analysis result of step 2;
[0019] 4. Use Cervus3.0 software to calculate the alle...
specific Embodiment approach 2
[0024] Embodiment two: the difference between this embodiment and embodiment one is: in step one, 9 pairs of microsatellite marker primers are divided into 4 groups to carry out multiple PCR reactions; the first group of PCR reactions consists of microsatellite marker 1, microsatellite marker 2 The primer pair of microsatellite marker 3 is carried out; the second group of PCR reactions is carried out by the primer pair of microsatellite marker 4 and microsatellite marker 5; the third group of PCR reactions is carried out by the primer pair of microsatellite marker 6 and microsatellite marker 7; The fourth set of PCR reactions was performed with primer pairs of microsatellite marker 8 and microsatellite marker 9 . Other steps and parameters are the same as those in Embodiment 1.
Embodiment 1
[0026] In 2013, 14 healthy males and 37 female broodstock were selected from the core population of Matsupu mirror carp in the Hulan Experimental Center of the Heilongjiang Fisheries Research Institute of the Chinese Academy of Fishery Sciences. Males mate with 2 to 3 females, and 37 full-sib families are established through artificial insemination. Collect fin ray samples of broodstock, record sex at the same time, and hatch fertilized eggs with different mating combination numbers separately. The offspring of Matsuura mirror carp were always cultured in the same pond and under the same conditions. At the 60th day of growth, all offspring of Matsuura mirror carp (1318 in total) were electronically marked and recorded, and fin rays were collected for genetic relationship identification.
[0027] 1. Extraction of individual genomic DNA from parents and offspring of Matsuura mirror carp:
[0028] (1) Take the fin ray tissue (about 60 mg per tail) of Matsuura mirror carp parent ...
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