| 松江鲈鱼ISSR-PCR反应体系的建立及优化研究(英文) |
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| 引用本文： | 郁建锋,鲍峰,韩晓磊,徐建荣.松江鲈鱼ISSR-PCR反应体系的建立及优化研究(英文)[J].农业科学与技术,2008,9(5):37-39. |
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| 作者姓名： | 郁建锋 鲍峰 韩晓磊 徐建荣 |
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| 作者单位： | 常熟理工学院生物与食品工程系 |
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| 基金项目： | 江苏省科技厅农业高技术项目
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| 摘 要： | Objective] The aim of this study is to establish and optimize the ISSR-PCR system in roughskin sculpin(Trachidermus fasciatus Heckel).Method] By using the primer of ISSR-63,the concentrations of Taq DNA polymerase,Mg2+,dNTPs and primer were optimized by orthogonal design for establishing the suitable ISSR-PCR system in roughskin sculpin;moreover,the suitable anneal temperature was yielded from gradient PCR on temperature.Result]The optimized ISSR-PCR system(20 μl reaction volume)in roughskin sculpin was proved to be:2.5 mmol/L Mg2+,250 μmol/L dNTPs,0.25 μmol/L primer,1 U Taq DNA polymerase,30 ng DNA template and 1×PCR buffer;and suitable anneal temperature was determined to be 50.8 ℃.The established system was further confirmed by using 24 wild roughskin sculpin samples.Conclusion]The results lay a foundation for the analysis of genetic diversity and germplasm resources of roughskin sculpin.
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| 关 键 词： | Trachidermus fasciatus Heckel Orthogonal design ISSR-PCR system |
Establishment and Optimization of ISSR-PCR System in Trachidermus fasciatus Heckel |
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| YU Jian-feng,BA Feng,HAN Xiao-lei,XU Jian-rong.Establishment and Optimization of ISSR-PCR System in Trachidermus fasciatus Heckel[J].Agricultural Science ＆ Technology,2008,9(5):37-39. |
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| Authors: | YU Jian-feng BA Feng HAN Xiao-lei XU Jian-rong |
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| Institution: | Department of Biology and Food Engineering,Changshu Institute of Technology,Changshu 215500 |
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| Abstract: | Objective] The aim of this study is to establish and optimize the ISSR-PCR system in roughskin sculpin (Trachidermus fasciatus Heckel). Method] By using the primer of ISSR-63, the concentrations of Taq DNA polymerase, Mg2+ , dNTPs and primer were optimized by orthogonal design for establishing the suitable ISSR-PCR system in roughskin sculpin; moreover, the suitable anneal temperature was yielded from gradient PCR on temperature. Result] The optimized ISSR-PCR system(20 μl reaction volume) in roughskin sculpin was proved to be: 2.5 mmol/L Mg2+, 250 μmol/L dNTPs, 0.25 μmoL/L primer, 1 U Taq DNA polymerase, 30 ng DNA template and 1×PCR buffer; and suitable anneal temperature was determined to be 50.8 ℃. The established system was further confirmed by using 24 wild roughskin sculpin samples. Conclusion] The results lay a foundation for the analysis of genetic diversity and germplasm resources of roughskin sculpin. |
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| Keywords: | Trachidermus fasciatus Heckel Orthogonal design ISSR-PCR system |
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