基于巢式PCR的重叠延伸PCR优化

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。%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.