文章摘要
聂鸿涛,李琪,孔令锋.皱纹盘鲍微卫星多重PCR体系构建及其在家系鉴定中的应用[J].水产学报,2013,37(2):207~215
皱纹盘鲍微卫星多重PCR体系构建及其在家系鉴定中的应用
Development of four multiplex PCR panels of microsatellites and application to kinship analysis in the Pacific abalone(Haliotis discus hannai)
投稿时间:2012-04-17  修订日期:2012-11-08
DOI:DOI:10.3724/SP.J.1231.2013.38112
中文关键词: 皱纹盘鲍  微卫星标记  多重PCR  家系鉴定
英文关键词: Haliotis discus hannai  microsatellites  multiplex PCR  parentage assignment
基金项目:国家自然科学基金项目(31072207);国家海洋局公益性行业科研专项(200905020,2012418023)
作者单位
聂鸿涛 大连海洋大学水产与生命学院,辽宁省贝类良种繁育工程技术研究中心 
李琪 中国海洋大学海水养殖教育部重点实验室 
孔令锋 中国海洋大学海水养殖教育部重点实验室 
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中文摘要:
      为提高微卫星分型效率,从以往报道的皱纹盘鲍微卫星中筛选出易扩增、特异性好的微卫星位点进行组合扩增,并通过优化退火温度、反应体系、引物浓度等条件,开发了4组多重PCR扩增体系。运用CERVUS 3.0 软件对12个皱纹盘鲍全同胞家系的372个子代进行家系鉴定,验证了这4组多重PCR在家系鉴定中的效率。结果发现,仅用1组微卫星多重PCR模拟和实际家系鉴定的成功率分别为86%和90%,两组则达到100%。结果表明,微卫星多重PCR技术能准确地把任意子代鉴定至其所属家系,可以进行大批量家系材料分析,具有较好的应用价值。
英文摘要:
      The Pacific abalone Haliotis discus hannai is a commercially important marine mollusk.To develop a rapid,economically efficient and robust approach for genetic studies in this species,twelve microsatellites were used to optimize four multiplex PCRs for the H.discus hannai,and the results obtained indicate the usefulness of the four multiplexed microsatellite systems in parentage assignment.We evaluated and validated these multiplex PCRs in 12 full-sib families.The average polymorphism information content(PIC)was 0.82.Inheritance analyses in the 144 tests showed that 4.9% microsatellite loci deviated from Mendelian ratios.The frequency of null alleles was estimated at 10.6% of all the alleles segregating based on a within-family analysis of Mendelian segregation patterns.Simulations and parentage analysis showed that with the most informative multiplex set 4,the assignment success could be 86% and 90% respectively,and 100% of the offspring were correctly allocated to their parents when two or more multiplex PCRs were used.Our results show that four multiplex PCR panels of microsatellites are possible and can be used for rapid and highly efficient parentage assignment in the Pacific abalone.
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