| 基于巢式PCR的重叠延伸PCR优化 |
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| 引用本文： | 彭雷,赵艳,马银花.基于巢式PCR的重叠延伸PCR优化[J].安徽农业科学,2016,44(20):126-127. |
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| 作者姓名： | 彭雷 赵艳 马银花 |
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| 作者单位： | 贵州师范大学生命科学学院,贵州贵阳,550001;武汉大学生命科学学院,湖北武汉,430072 |
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| 基金项目： | 国家自然科学基金项目“水稻抗褐飞虱基因多样性持续控制褐飞虱的分子机理”(31230060)。 |
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| 摘 要： | 目的]结合巢式PCR对重叠延伸PCR进行优化。方法]以褐飞虱Actin 1基因启动子区与EGFP表达盒区基因融合为例,通过巢式PCR对重叠延伸PCR进行优化。结果]成功获得融合片段,经过巢式PCR优化后大大简化试验流程,缩短试验时间,并增强PCR特异性与检出率。结论]优化了重叠延伸PCR,可更快速高效融合基因片段。
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| 关 键 词： | 重叠延伸PCR 巢式PCR 褐飞虱 Actin 1启动子 |
Optimization of Splicing by Overlap Extension PCR Using Nested PCR |
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| PENG Lei,ZHAO Yan,MAYin-hua.Optimization of Splicing by Overlap Extension PCR Using Nested PCR[J].Journal of Anhui Agricultural Sciences,2016,44(20):126-127. |
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| Authors: | PENG Lei ZHAO Yan MAYin-hua |
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| Abstract: | Objective] To optimize the Splicing by overlap extension PCR using nested PCR.Method] Nested PCR was used to optimize SOE-PCR (splicing by overlap extension PCR) for combining brown planthopper (BPH) Actin 1 promoter and EGFP expression cassette. Result] Nested PCR could greatly optimize SOE-PCR.After optimization of nested PCR, the test process was simplified, and the detection time was shortened.The PCR specificity and detection rate were enhanced. Conclusion] The optimized SOE-PCR can rapidly and effectively fuse the gene segment. |
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| Keywords: | SOE-PCR Nested PCR Brown planthopper Actin 1 promoter |
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