Cassette mutagenesis is a type of site-directed mutagenesis that uses a short, double-stranded oligonucleotide sequence (gene cassette) to replace a fragment of target DNA. It uses complementary restriction enzyme digest ends on the target DNA and gene cassette to achieve specificity. It is different from methods that use single oligonucleotide in that a single gene cassette can contain multiple mutations. Unlike many site directed mutagenesis methods, cassette mutagenesis also does not involve primer extension by DNA polymerase.[1][2][3][4][5]
^Worrall, Andrew (1994). "Site-Directed Mutagenesis by the Cassette Method". Methods in Molecular Biology. Vol. 30. Humana Press. pp. 199–210. doi:10.1385/0-89603-256-6:199. ISBN 978-1-59259-517-4. PMID 8004195.
^Wells, J. A.; Vasser, M; Powers, D. B. (1985). "Cassette mutagenesis: An efficient method for generation of multiple mutations at defined sites". Gene. 34 (2–3): 315–23. doi:10.1016/0378-1119(85)90140-4. PMID 3891521.
^Clore, Adam; Reinertson, Brian; Rose, Scott; Sabel, Jaime. "Ultramer Oligonucleotides Mutagenesis Application Guide - Experimental Overview, Protocol, Troubleshooting" (PDF). WWW.IDTDNA.COM. Integrated DNA Technologies. p. 16. Archived from the original (PDF) on December 5, 2014. Retrieved Nov 6, 2014.
^Kegler-Ebo, D. M.; Docktor, C. M.; Dimaio, D (1994). "Codon cassette mutagenesis: A general method to insert or replace individual codons by using universal mutagenic cassettes". Nucleic Acids Research. 22 (9): 1593–9. doi:10.1093/nar/22.9.1593. PMC 308034. PMID 8202358.
^El-Mansi, E. M. T.; Bryce, C. F. A.; Demain, Arnold L.; A.R. Allman (2006-10-25). Fermentation Microbiology and Biotechnology, Second Edition. CRC Press. pp. 222–. ISBN 978-0-8493-5334-5. Retrieved 27 November 2014.
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