A microsatellite marker linked to high fecundity traits of Procambarus clarkii and its application
A technology of microsatellite markers and Procambarus clarkii, applied in the field of animal molecular genetics and breeding, to speed up the progress of breeding, overcome low efficiency and reduce investment
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Embodiment 1
[0029] 1. Take 101 Procambarus clarkii with eggs from the Procambarus clarkii farm in Yancheng City, Jiangsu Province, weigh the weight of each Procambarus clarkii, count the amount of eggs held by each Procambarus clarkii, and calculate the eggs of each Procambarus clarkii The ratio of number to weight. Sorting according to the ratio results, take the 24 individuals with the smallest relative egg holding capacity to form a low-fecundity group. A high-fecundity group is formed, and the value of the relative egg mass of each individual of the group is greater than 10 (egg grain / gram body weight). The tissue of Procambarus clarkii was collected, and the DNA of Procambarus clarkii was extracted, and the extraction method adopted the traditional phenol-chloroform extraction method. Specific steps are as follows:
[0030] 1.1 Put 0.1g muscle from each individual into a 1.5ml centrifuge tube, cut it into pieces, add 450μL STE extraction buffer (10mmol / L Tris-HCl, pH8.0; 1mmol / L ED...
Embodiment 2
[0043] 1. In October 2016, 2,000 Procambarus clarkii parents with eggs were collected from a Procambarus clarkii farm in Yancheng City, Jiangsu Province. The individuals were healthy, growing well, with complete appendages. Use a tightenable plastic collar with a numbered number to buckle at the base of the claws of the Procambarus clarkii to number each brooding shrimp;
[0044] 2. For the fertilized eggs of Procambarus clarkii attached to the appendages of the mother's abdomen at different developmental stages, use sterile tweezers to pick up 5-10 fertilized eggs (a total of 15-30 eggs) in the front, middle and back of the abdomen. In eppendorf tubes, quick-frozen in liquid nitrogen and brought back to the laboratory for temporary storage in a -80°C refrigerator;
[0045] 3. The genomic DNA of the embryos of Procambarus clarkii 2000 at different developmental stages was extracted using Soil Genomic DNA Kit from Beijing Kangwei Century Bioreagent Co., Ltd.
[0046] 4. Adopt ...
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