Abstract
Targeted nucleotide exchange (TNE) is a process in which an oligonucleotide bearing sequence complementarity aligns with the sequence of a target gene and directs the alteration of a single base. This technique can be used to repair a point mutation or mediate site-specific mutagenesis. A critical factor in the development of this approach centers around the elevation and stabilization of the frequencies with which these events occur. Here we describe a protocol for increasing the frequency of TNE in the true yeast, Saccharomyces cerevisiae, through the use of nonspecific, carrier oligonucleotides. These molecules, when added to the reaction, increase the TNE frequency up to 25-fold in some cases, perhaps by providing a molecular trap to bind factors, which may inactivate the specific targeting oligos.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
