Functional genomics requires vector construction for protein expression and functional characterization of target genes; therefore, a simple, flexible, and low-cost molecular manipulation strategy will be highly advantageous for genomics approaches. Here, we describe a ohm-PCR strategy that enables multiple types of sequence modification, including precise insertion, deletion and substitution, in any position of a circular plasmid. ohm-PCR is based on an overlap extension site-directed mutagenesis technique, and is named for its characteristic ohm-shaped secondary structure during PCR. ohm-PCR can be performed either in two steps, or in one tube in combination with exonuclease I treatment. These strategies have wide applications for protein engineering, gene function analysis and in vitro gene splicing.
Robust one-tube ohm-PCR strategy accelerates precise sequence modification of plasmids for functional genomics.: "Publication Date: 2013 Jan 17 PMID: 23335613
Authors: Chen, L. - Wang, F. - Wang, X. - Liu, Y. G.
Journal: Plant Cell Physiol
(Via Plant and Cell Physiology.)