| 青鱼ISSR-PCR反应体系的建立及优化研究 |
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| 引用本文： | 张建铭,吴志强,胡茂林,曾庆祥.青鱼ISSR-PCR反应体系的建立及优化研究[J].福建水产,2010(4):12-15. |
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| 作者姓名： | 张建铭 吴志强 胡茂林 曾庆祥 |
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| 作者单位： | [1]江西省赣州市水产研究所,江西赣州341000 [2]桂林理工大学环境科学与工程学院,广西桂林541001 [3]南昌大学生命科学与食品工程学院,江西南昌330031 |
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| 摘 要： | 以赣江野生青鱼(Mylopharyngodon piceus)基因组DNA为试验材料,研究Mg2+浓度、dNTP浓度、引物浓度和模板浓度4个参数对ISSR-PCR反应的影响,建立一套适合青鱼ISSR-PCR反应的最佳体系。结果表明,在25μL的反应体系中,Mg2+、dNTP、引物和模板4个参数的最适宜浓度分别为:3.0mmol/L、0.25mmol/L、0.4umol/L和30ng。PCR扩增程序为:94℃预变性5min,94℃30s,52℃45s,72℃2min,共38个循环,最后72℃继续延伸10min。
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| 关 键 词： | 青鱼 ISSR 反应体系 |
Establishment and optimization of ISSR-PCR system in Mylopharyngodon piceus |
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| ZHANG Jian-ming,WU Zhi-qiang,HU Mao-lin,ZENG Qing-xiang.Establishment and optimization of ISSR-PCR system in Mylopharyngodon piceus[J].Journal of Fujian Fisheries,2010(4):12-15. |
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| Authors: | ZHANG Jian-ming WU Zhi-qiang HU Mao-lin ZENG Qing-xiang |
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| Institution: | 1.Ganzhou fisheries research institute,Ganzhou,341000,China;2.School of Environmental Sciences and engineering Guilin University of technology,Guilin 541001,China;3.School of Life Sciences and food engineering,Nanchang University,Nanchang,330031,China.) |
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| Abstract: | In this study,the wild black carp＇s（Mylopharyngodon piceus） genomic DNA of Ganjiang river was used as the experimental material.An ISSR-PCR reaction system was constructed by the optimization of the concentrations of Mg2＋,dNTPs,primer and template.The results showed that the conditions for ISSR-PCR system of black carp were as follows： PCR reaction volume 25μL,including 3.0mmol/L Mg2＋,0.25mmol/L dNTPs,0.4umol/L primer and 30ng template.The PCR reaction program was with 5 minutes at 94℃,which is followed 38 cycles,each with 30 seconds at 94℃,45 seconds at 52℃,2 minutes at 72℃,and a final extension at 72℃ for 10 minutes. |
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| Keywords: | Mylopharyngodon piceus ISSR reaction condition |
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