| Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis |
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| 作者姓名： | GAO Huan KONG Jie LIU Ping MENG Xianhong LUAN Sheng ZHANG Tianshi |
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| 作者单位： | GAO Huan1，2，KONG Jie2，LIU Ping2，MENG Xianhong2，LUAN Sheng2，ZHANG Tianshi21. Jiangsu Key Laboratory of Marine Biotechnology，Huaihai Institute of Technology，Lianyungang 222005，China2. Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture，Yellow Sea Fisheries Research Institute，Chinese Academy of Fishery Sciences，Qingdao 266071，China |
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| 基金项目： | 国家高技术研究发展计划(863计划)，引进国际先进农业科技计划(948计划)，国家自然科学基金，the Open-end Funds of Jiangsu Key Laboratory of Manne Biotechnology，the Huaihai Institute of Technology，the Open-end Funds of Key Laboratory of Aquatic Genetic Resources and Aquacultural Ecosystem of the Ministry of Agriculture of China，Shanghai Fisheries University |
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| 摘 要： | Through exploring the microsatellite primers from the random genome sequences of Chinese shrimp (Fenneropenaeus chinensis), some microsatellite primers were obtained with rich polymorphic genetic information, and a triplex PCR was established using three primers (RS1101, RS0683 and H081 primers). By adjusting the final concentration of Mg2 , dNTP and primers, and using a touch-town PCR program, the optimum amplification parameters of PCR system were obtained, which could successfully amplify the three primers in a PCR reaction. In the denatured PAGE gel, the amplified DNA fragments of three primers RS1101, RS0683 and H081 could be easily identified each other. For the triplex PCR system, the PPE (probabilities of paternity exclusion) is 0.967 9, and the DP (discrimination power) is 0.999 327. Using the triplex PCR to test ten individuals of a parentage and their parents, an individual was excluded from the parentage in all of the three microsatellite loci, which might be mixed into the parentage for some unknown reason such as factitious misplay. The triplex PCR will be of great practical value in identifying the parentages of F. chinensis.
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| 关 键 词： | 中国明对虾 微卫星 亲本分析 三重PCR |
| 收稿时间： | 2006-02-01 |
| 修稿时间： | 2006-12-31 |
Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis |
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| GAO Huan,KONG Jie,LIU Ping,MENG Xianhong,LUAN Sheng,ZHANG Tianshi.Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis[J].Acta Oceanologica Sinica,2007,26(2):65-74. |
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| Authors: | GAO Huan KONG Jie LIU Ping MENG Xianhong LUAN Sheng and ZHANG Tianshi |
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| Institution: | Jiangsu Key Laboratory of Marine Biotechnology, Huaihai Institute of Technology, Lianyungang 222005, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China |
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| Abstract: | Through exploring the microsatellite primers from the random genome sequences of Chinese shrimp (Fenneropenaeus chinensis), some microsatellite primers were obtained with rich polymorphic genetic information, and a triplex PCR was established using three primers (RS1101, RS0683 and H081 primers). By adjusting the final concentration of Mg2 , dNTP and primers, and using a touch-town PCR program, the optimum amplification parameters of PCR system were obtained, which could successfully amplify the three primers in a PCR reaction. In the denatured PAGE gel, the amplified DNA fragments of three primers RS1101, RS0683 and H081 could be easily identified each other. For the triplex PCR system, the PPE (probabilities of paternity exclusion) is 0.967 9, and the DP (discrimination power) is 0.999 327. Using the triplex PCR to test ten individuals of a parentage and their parents, an individual was excluded from the parentage in all of the three microsatellite loci, which might be mixed into the parentage for some unknown reason such as factitious misplay. The triplex PCR will be of great practical value in identifying the parentages of F. chinensis. |
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| Keywords: | multiplex PCR touch-down PCR microsatellite parentage analysis Fenneropenaeus chinensis |
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