Development and Characteristics of Tetranucleotide Repeat Microsatellite Loci in Hyriopsis cumingii

Abstract

Abstract: A microsatellite-enriched library of the Hyriopsis cumingii was constructed using repeat-enrichment method with biotin-labeled oligo（ATAC）₅ and streptavidin coated magnetic beads. Forty-three pairs of primers designed by software PrimerSelect according to the sequences were synthesized for the pre-screening. Twenty pairs of primers which could amplify constant and legible expected DNA products were labeled by fluorescence. Seventeen microsatellite loci showed high levels in genetic polymorphism testing on 36 individuals sampled from Dongting Lake. The number of alleles at each locus ranged from 6 to 28. The expected and observed heterozygosities varied from 0.677 6 to 0.946 4 and from 0.638 9 to 1.000, respectively. The PIC value ranged from 0.630 to 0.929. Twelve microsatellite loci fitted to Hardy-Weinberg equilibrium. Compared with the previous dinucleotide repeat microsatellite developed in our laboratory, these 17 tetranucleotide repeat microsatellite loci showed higher polymorphism. Also, these 17 microsatellite loci can make genotyping easier by conventional methods such as polyacrylamide gel electrophoresis and save cost. Therefore, it offered accurate and inexpensive microsatellite markers for population genetic analysis and paternity test in Hyriopsis cumingii.

Key words: Hyriopsis cumingii, Microsatellite, Tetranucleotide repeat, Polymophism

Han Xuekai,Li Jiale,Wang Zhaoqi,Bai Zhiyi. Development and Characteristics of Tetranucleotide Repeat Microsatellite Loci in Hyriopsis cumingii[J]. Biotechnology Bulletin, 2014, 0(6): 139-144.

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