| An Efficient Procedure for Isolating Microsatellite DNAs from Sea Cucumber(Apostichopus japonicus) |
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| 作者姓名： | HU Jingjie ZHAN Aibin LU Wei HU Xiaoli and BAO Zhenmin* Laboratory of Marine Genetics and Breeding |
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| 作者单位： | HU Jingjie（Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China） ;
ZHAN Aibin（Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China） ;
LU Wei（Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China） ;
HU Xiaoli（Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China） ;
BAO Zhenmin（Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China） ; |
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| 基金项目： | 国家高技术研究发展计划(863计划);国家自然科学基金 |
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| 摘 要： | 1 Introduction Microsatellite DNAs, also called simple sequence re-peats (SSRs), are stretches of DNA, consisting of tan-demly repeating mono-, di-, tri-, tetra- or penta-nucleotide units. They are evenly distributed throughout the ge-nomes of most eukaryotic species (Tohn et al., 2000). Their high information content about the divergence of genome, which is directly related to the number of alleles at each locus, and the ease of PCR assays make them ex-cellent molecular markers (Powell …
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An Efficient Procedure for Isolating Microsatellite DNAs from Sea Cucumber (Apostichopus japonicus) |
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| HU Jingjie,ZHAN Aibin,LU Wei,HU Xiaoli,and BAO Zhenmin* Laboratory of Marine Genetics and Breeding.An Efficient Procedure for Isolating Microsatellite DNAs from Sea Cucumber(Apostichopus japonicus)[J].Journal of Ocean University of China,2007,6(1):12-15. |
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| Authors: | HU Jingjie ZHAN Aibin LU Wei HU Xiaoli BAO Zhenmin |
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| Institution: | Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Biotechnology,Ocean University of China, Qingdao 266003, P. R. China |
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| Abstract: | The construction of enrichment library proves to be one of the efficient approaches for isolating microsatellites in this study. The genomic DNA of sea cucumber was digested with HaeIII and size-selected DNA fragments (250-700 bp) were ligated to an adaptor. Microsatellite-containing sequences were captured by using a combination of GA and CA probes, which were attached to a nylon membrane. The microsatellite enrichment library constructed in this study consisted of approximately 700 clones. Two hun-dred and thirty-two clones reacted positively after the library screening procedure. Of the 50 clones sequenced, all contained at least one microsatellite and one duplicate clone was found. Approximately 86% of the sequenced fragments permitted to design primers for sequence tagged microsatellite site (STMS). |
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| Keywords: | SSRs enrichment sea cucumber Apostichopus japonicus |
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