Splicing together different regions of a gene by modified polymerase chain reaction-based site-directed mutagenesis.
A modified polymerase chain reaction (PCR)-based site-directed mutagenesis method used to splice together different regions of a gene by deleting hundreds of nucleotides of undesired sequences is described. This method was inspired by a PCR-based site-directed mutagenesis method developed by Stratagene (La Jolla, CA, USA); the procedure and primer design were modified to enable the method to generate deletions several hundreds of nucleotides in length with an efficiency of 80-100%, and to delete two DNA fragments simultaneously in a single PCR. This method should be useful for deletion of large DNA fragments from a gene.
Li
Xiangdong
X
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai, 200232, People's Republic of China.
Qiu
Yafeng
Y
Shen
Yang
Y
Ding
Chan
C
Liu
Peihong
P
Zhou
Jinping
J
Ma
Zhiyong
Z
D000818:Animals; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D017353:Gene Deletion; D006801:Humans; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004247:DNA