| 转基因玉米PCR检测体系的优化研究 |
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| 引用本文： | 何春梅,张成华,刘强,董瑞,高新学,汪黎明,刘铁山.转基因玉米PCR检测体系的优化研究[J].山东农业科学,2011(6):14-16,19. |
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| 作者姓名： | 何春梅 张成华 刘强 董瑞 高新学 汪黎明 刘铁山 |
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| 作者单位： | 山东省农业科学院玉米研究所,山东济南,250100 |
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| 基金项目： | 国家科技支撑计划项目，山东省良种工程项目资助 |
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| 摘 要： | 以转基因玉米为研究材料,从模板DNA的提取质量、Mg2＋浓度、退火温度、引物用量等方面对PCR检测体系进行优化,建立了检测转基因玉米中外源基因的有效PCR反应体系。在优化体系中,25mm ol/LMg2＋用量由1.5μl降低为1.2μl,12.5mm ol/L引物用量由1μl降低为0.5μl,退火温度提高2℃。TouchDown PCR反应条件：95℃5m in;95℃1m in,62.5℃1m in（以后每个循环递减1℃）,72℃1m in,循环15次;95℃1m in,57.5℃1m in,72℃1m in,循环20次;72℃延伸7m in。
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| 关 键 词： | 转基因玉米 PCR扩增 反应体系 |
Optimization of PCR System for Transgenic Maize |
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| HE Chun-mei,ZHANG Cheng-hua,LIU Qiang,DONG Rui,GAO Xin-xue,WANG Li-ming,LIU Tie-shan.Optimization of PCR System for Transgenic Maize[J].Shandong Agricultural Sciences,2011(6):14-16,19. |
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| Authors: | HE Chun-mei ZHANG Cheng-hua LIU Qiang DONG Rui GAO Xin-xue WANG Li-ming LIU Tie-shan |
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| Institution: | (Maize Institute,Shandong Academy of Agricultural Sciences,Jinan 250100,China) |
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| Abstract: | With the transgenic maize as experiment material,the PCR system for detecting exogenous gene was optimized in extracted DNA quality,Mg2+ concentration,annealing temperature and primer dosage.In the optimized system,the dosage of 25 mmol/L Mg2+ was reduced from 1.5 μl to 1.2 μl;the dosage of 12.5 mmol/L primer was reduced from 1 μl to 0.5 μl;the annealing temperature was increased by 2℃.The Touchdown PCR reaction conditions were 95℃ for 5 min;95℃ for 1 min,62.5℃ for 1 min(1℃ decreased after each cycle),72℃ for 1 min,15 cycles;95℃ for 1 min,57.5℃ for 1 min,72℃ for 1 min,20 cycles;and 72℃ for 7 min for final extension. |
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| Keywords: | Transgenic maize PCR amplification Reaction system |
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