Two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof
A technology of microsatellite markers and large yellow croaker, applied in biochemical equipment and methods, DNA preparation, microbial measurement/inspection, etc., can solve the problems of lack of systematic research, shorten the breeding process and improve the efficiency of breeding
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Embodiment 1
[0015] Example 1: Screening method for fast and long microsatellite markers
[0016] The large yellow croaker material used is a full-sibling family (referred to as FS) bred by artificial insemination in an experimental base in Fujian Province. 208 large yellow croakers were collected from FS, and the heaviest and lightest large yellow croakers were selected based on body weight. There were 30 large yellow croakers, and the body length, height and weight of each large yellow croaker were measured. Part of the caudal fin was cut and fixed in 95% ethanol for DNA extraction.
[0017] DNA extraction and detection Use conventional phenol / chloroform / isoamyl alcohol method to extract DNA, UV spectrophotometer to measure OD 260 with OD 280 , determine the quality of the DNA and adjust the DNA concentration to 50ng / μL, and store it at -20°C for later use.
[0018] PCR and detection PCR reaction system: the total reaction volume is 20μL, including 1.0μL (50ng) template DNA, 2.0μL 10×...
Embodiment 2
[0030] Example 2: Preliminary verification of microsatellite markers
[0031] In order to verify the effectiveness of the screening markers, another 60 families were selected from the above-mentioned families for fast and long marker screening for verification analysis, and the experimental procedure was as described in Example 1. This time, only the relevant sites in the preliminary screening were analyzed, namely LYC0088, LYC0143 and LYC0200, and the primer sequences used were SEQUENCE LISTING1-SEQUENCELISTING6 in the sequence list. It has been verified that there are two loci related to growth traits, namely loci LYC0088 and LYC0143. These two loci are significantly correlated with the length, height and weight of large yellow croaker (P<0.01). The fast-growing markers of large yellow croaker are preliminary The verification results are shown in Table 2.
[0032] Table 2
[0033]
Embodiment 3
[0034] Example 3: Revalidation of microsatellite markers
[0035] The 2 microsatellite markers screened were used to genotype the largest 30 fish and the smallest 30 fish in another large yellow croaker population (the progeny of mixed random mating of 4 females and 4 males), and a total of 60 individuals were combined. The phenotypic data of the three traits of body length, body height, and body weight of each individual were tested for significance, and the results of the re-verification of the fast-length marker of large yellow croaker can be seen in Table 3.
[0036] table 3
[0037]
[0038] It can be seen from the test results that LYC0088 and LYC0143 are still closely related to growth traits, among which LYC0088 has a very significant correlation with body height and weight (P<0.01), and a significant correlation with body length (P<0.05), and LYC0143 has a significant correlation with body length and body weight. The correlation between body weight was significant...
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