| 摘要: |
| 筛选多态性高、特异性好、片段大小适中的10 个长牡蛎微卫星位点进行优化组合, 根据扩 增片段大小及同一荧光不重叠原则构建了两组五重PCR 体系。运用CERVUS 3.0 软件对0527 组27 个长牡蛎全同胞家系的643 个子代和0612 组27 个全同胞家系382 个子代分别进行亲权鉴定。结果 发现, 用两组微卫星五重PCR 鉴别时, 在0527 组和0612 组的鉴定成功率均为100%; 只用第一组微 卫星五重PCR, 可以将0527 组96%的子代和0612 组96%的子代鉴定到亲本; 只用第二组微卫星五 重PCR, 可以将0527 组97%的子代和0612 组95%的子代鉴定到亲本。本研究中筛选出的两组微卫 星五重PCR 体系在两组家系中的鉴定效率均较高, 可以快速有效地将子代个体鉴定至所属父母本, 在长牡蛎家系鉴定中具有很好的应用价值。 |
| 关键词: 长牡蛎 微卫星 五重PCR 亲子鉴定 |
| DOI：10.11693/hyhz20150900231 |
| 分类号: |
| 基金项目:国家自然科学基金青年项目, 31402298 号; 山东省农业良种工程项目--大宗经济贝类新品种选育及应用; 黄河三角洲 学者--海洋生物遗传育种岗位; 烟台市科技发展计划项目, 2015ZH073 号。 |
附件 |
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| DEVELOPMENT OF MULTIPLEX PCR OF MICROSATELLITES AND ITS APPLICATION TO KINSHIP ANALYSIS IN CRASSOSTREA GIGAS |
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JI Ren-Ping1,2, LI Huan-Jun2, FENG Yan-Wei2, ZHANG Rong-Liang1,2, WANG Wei-Jun2, YANG Jian-Min2
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1.College of Fishes and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.Shandong Marine Resource and Environment Research Institute, Shandong Provincial Key Laboratory of Restoration for Marine Ecological Restoration, Yantai 264006, China
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| Abstract: |
| We tested two sets of high-level 5-multiplex PCRs for efficient genotyping in the Pacific oyster (Crassostrea gigas). Ten loci were selected based on polymorphism, reliability, and allele size range. They were divided into two groups according to allele size and labeled with four different fluorescence dyes, so that loci in the same color were not be overlap in size. In addition, we performed a paternity test for 27 full-sib families and 643 offspring of Group 0527 and 27 full-sib families and 382 offspring of Group 0612 with software CERVUS 3.0. Parentage analysis demonstrated that 100% of all offspring were allocated unambiguously to the unique pair of parents when two sets of 5-multiplex PCRs were used for Groups 0527 and 0612. The assignment success for the two groups was 96% based on MS 1 (the first 5-multiplex PCR). However, the assignment success was 97% and 95% based on MS 2 (the second 5-multiplex PCR) for Groups 0527 and 0612 respectively. Therefore, the two sets of 5-multiplex PCRs can be used for rapid and efficient parentage assignment in the Pacific oyster. |
| Key words: Crassostrea gigas microsatellite multiplex PCR parentage analysis |
