An efficient approach for site-directed mutagenesis using central overlapping primers
Date
2011-07-18Author
Wang, Huiping
Zhou, Nan
Ding, Feng
丁凤
Li, Zhaofang
Chen, Ruichuan
陈瑞川
Han, Aidong
韩爱东
Liu, Runzhong
Collections
- 生命科学-已发表论文 [5901]
Abstract
QuikChange is a popular method for site-directed mutagenesis in structural and functional studies of proteins and nucleic acids. However, the standard protocol is often inefficient in producing the desired mutations. Here we present a novel strategy for primer design, central overlapping primers (COP), which employs a pair of bipartite primers of different lengths, with the short primer complementary to the middle region of the long primer. The COP method is efficient and robust in generating approximately 90% mutation rate without supercompetent Escherichia coli cells or laborious screening for positive clones. (C) 2011 Elsevier Inc. All rights reserved.
Citation
Anal Biochem. 2011 Nov 15;418(2):304-6.URI
http://dx.doi.org/doi:10.1016/j.ab.2011.07.008WOS:000295240100020
https://dspace.xmu.edu.cn/handle/2288/11776