Selection And Application Of SSR Markers Based On Sika Deer Genome

China is the earliest country for sika deer breeding.Deer resources are very rich,but wrong pedigree records are common during sika deer breeding.Microsatellite primers are developed based on the preliminary completion of the sika deer genome sequencing,and molecular deer sika deer molecular markers are used.Provide a theoretical basis for breeding and breeding.1.Based on de novo sequencing of sika deer genomes and weight sequencing of 100 deer sikas,the SSR locus in the sika deer genome was analyzed using MISA,the SSR locus of 4 base repeat units was selected,the SSR interval of InDels was extracted,and Primer3 pairs were selected.The SSR locus was designed in batches and primers were used for e-PCR to remove non-specific amplification.Based on the position of the amplified fragment in the bovine chromosome,the selected SSR markers should cover all chromosomes as much as possible,avoiding the end of the chromosome.100 pairs of primers were used to perform polyacrylamide gel electrophoresis on 8 sika individuals.31 pairs of microsatellite primers were screened and polymorphisms were verified.The minimum PIC was 0.572.Generally,PIC>0.5 was considered to be highly polymorphic.For subsequent group diversity analysis and paternity testing.2.A total of 31 pairs of microsatellite primers were used to analyze the polymorphisms of four populations of sika deer.Capillary electrophoresis was used for genotyping.A total of 434 alleles were found in the four sika populations,and the average expected heterozygosity and observation were mixed.The merging degrees were 0.6180 and 0.5254 respectively.The PICs of the four populations of Dongda,Siping,Shuangyang and Dongfeng were 0.5804,0.5428,0.5715 and 0.5783 respectively.The mean population PIC was 0.5683,indicating that the sika deer population was rich in genetic diversity.By analysis,the genetic differences between the populations ranged from 0.092(C vs D)to 0.158(A vs D),and the Nei's genetic distance was used to examine the differences between populations and between populations to explain the genetic differences and differences among the four populations.The results showed that the genetic differences between groups and groups were not significant.3.31 pairs of microsatellites were used to amplify sika deer.The paternity analysis was performed by comparison of the peak maps and cervus software.The cumulative exclusion rate of 31 pairs of primers was more than 99.99% in all three modes.The average parental exclusion rate was 0.6714 ? 0.5134 and 0.3429,and 11 pairs of Dongdaluchang,2 pairs of Shuangyangluchang,4 pairs of Sipingluchang,and 17 pairs of father and son in Zuojialuchang and 5 pairs of mother and son in Zuojialuchang were determined.Exclude candidate parent individuals SY-46(SY-198),SY-12(SY-46),SY-98(SY-79),SY-100(SY-507),SY-307(SY-098,SY)-598)and maternal individual M48(Z018).