A Modified Method of QuikChange Site-Directed Mutagenesis
- OAPS－大学生创新计划项目 
[中文文摘]DNA传统的快速点突变方法即"一步突变法"虽然可使DNA产生突变,但效率不高.市场上出售的快速点突变试剂盒,其突变效率可达80%以上,然而价钱昂贵.本文论述的快速点突变方法,在传统的"一步突变法"基础上做了改进,通过对引物突变点5′端序列Tm值的确定,使引物设计变得简单,初学者可轻松完成DNA的定点突变,而且大大降低了成本,适用于对大量DNA进行定点突变试验.试验结果表明:这种经过改良的方法突变率为96.22%,可为研究人员的试验带来便利,是一种值得推广的新方法.[英文文摘] traditional method that introduced mutations to DNA was the QuikChangeTM Site-Directed Mutagenesis,called"One-Step Site-Directed Mutagenesis".However,it had low efficiency.The QuikChangeTM Site-Directed Mutagenesis Kit could create mutants about 80% efficiency,but it was too expensive.The protocol in this paper was a modification of the traditional "One-Step Site-Directed Mutagenesis" and simplified the design of primers so that the DNA could be easily mutated by novice.Our results indicated that the efficiency of this modified method could reach 96. 22% .Th is protocolm aight be applied for a large number of DNA samples needed to bem utated due to its low co s.t So it is worth widely application.