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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,(2):65-74
Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis
Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis
投稿时间:2006-02-01  修订日期:2006-12-31
DOI:
中文关键词:  multiplex PCR  touch-down PCR  microsatellite  parentage analysis  Fenneropenaeus chinensis
英文关键词:multiplex PCR  touch-down PCR  microsatellite  parentage analysis  Fenneropenaeus chinensis
基金项目:
作者单位E-mail
GAO Huan 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 
 
KONG Jie 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 kongjie@sina.com 
LIU Ping 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  
MENG Xianhong 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  
LUAN Sheng 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  
ZHANG Tianshi 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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中文摘要:
      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.
英文摘要:
      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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