Journal
BIOTECHNOLOGY LETTERS
Volume 29, Issue 6, Pages 925-930Publisher
SPRINGER
DOI: 10.1007/s10529-007-9327-4
Keywords
asymmetric overlap extension PCR; asymmetric PCR; overlap extension PCR; site-directed mutagenesis
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By combining asymmetric PCR and overlap extension, we developed a novel asymmetric overlap extension PCR (AOE-PCR) method for site-directed mutagenesis which bypassed the need for intermediate purification and excluded the amplification of a wild-type template. This method was used to introduce single base mutations into a small GTPase gene from cotton and to simultaneously introduce two mutations just by repeating this method using the first round AOE-PCR products as template. Our results suggested that the AOE-PCR method represents a valuable improvement of the original overlap extension PCR for site-directed mutagenesis.
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