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An improved dual-tube megaprimer approach for multi-site saturation mutagenesis
Stockholm University, Faculty of Science, Department of Neurochemistry.ORCID iD: 0000-0002-6416-064X
2013 (English)In: World Journal of Microbiology & Biotechnology, ISSN 0959-3993, E-ISSN 1573-0972, Vol. 29, no 4, 667-672 p.Article in journal (Refereed) Published
Abstract [en]

Saturation mutagenesis is a powerful tool in protein engineering. Even though QuikChange site-directed mutagenesis method is dominantly used in laboratories, it could not be successfully applied to the generation of a focused mutant library of human glutathione transferase A2-2. In the present study, we further developed an improved versatile dual-tube approach of randomizing difficult-to-amplify targets, exhibiting significant improvement towards equal distribution of nucleotides at randomized sites compared to other published methods.

Place, publisher, year, edition, pages
2013. Vol. 29, no 4, 667-672 p.
Keyword [en]
Protein engineering, Saturation mutagenesis, Megaprimer approach, QuikChange method
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:su:diva-89567DOI: 10.1007/s11274-012-1222-zISI: 000316291000010OAI: oai:DiVA.org:su-89567DiVA: diva2:618880
Funder
Swedish Research CouncilSwedish Cancer Society
Note

AuthorCount:2;

Available from: 2013-04-30 Created: 2013-04-29 Last updated: 2017-12-06Bibliographically approved

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Mannervik, Bengt
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