基于巢式PCR的重叠延伸PCR优化
Optimization of Splicing by Overlap Extension PCR Using Nested PCR
彭雷 1赵艳 2马银花2
作者信息
- 1. 贵州师范大学生命科学学院，贵州贵阳550001
- 2. 武汉大学生命科学学院，湖北武汉430072
- 折叠
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。
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.
关键词
重叠延伸PCR/巢式PCR/褐飞虱/Actin/1启动子Key words
SOE-PCR/Nested PCR/Brown planthopper/Actin 1 promoter引用本文复制引用
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据