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Updated: Jul 10, 2026

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
Published on: February 5, 2019
Splicing together different regions of a gene by modified polymerase chain reaction-based site-directed mutagenesis
Xiangdong Li1, Yafeng Qiu, Yang Shen
1Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai, 200232, People's Republic of China.
Abstract:
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.
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