Abstract
A method for random mutagenesis using primers synthesized from four phosphoramidite solutiosn that have been “spiked” with a small amount of the other three residues is presented. The random oligonucleotides are used in an improved phosphorothioate mutagenesis technique that allows the entire procedure, including transformations, to be completed in 1 day. The phosphorothioate technique gives a high efficiency of mutagenesis from oligonucleotides with random mutations. A large proportion of the transformants analyzed by sequencing are found to contain random changes. The spiked oligonucleotides give truly random mutagenesis within the sequence defined by the primer. This is a very effective method for determining the functionally important amino acids in a protein when little information apart from the DNA sequence is known.
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.
