| GAO Huan,KONG Jie,LIU Ping,MENG Xianhong,LUAN Sheng,ZHANG Tianshi. 2007. Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis. Acta Oceanologica Sinica, (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 |
| Received:February 01, 2006 Revised:December 31, 2006 |
| DOI： |
| Key words:multiplex PCR touch-down PCR microsatellite parentage analysis Fenneropenaeus chinensis |
| 中文关键词: multiplex PCR touch-down PCR microsatellite parentage analysis Fenneropenaeus chinensis |
| 基金项目: |
| Author Name | Affiliation | 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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| 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. |
| 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. |
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