An improved dual-tube megaprimer approach for multi-site saturation mutagenesis

  • Wei Zhang
  • Bengt MannervikEmail author
Original Paper


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.


Protein engineering Saturation mutagenesis Megaprimer approach QuikChange method 



QuikChange site-directed mutagenesis method


Glutathione transferase


Polymerase chain reaction



This work was supported by grants from the Swedish Research Council and the Swedish Cancer Society.


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Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.Department of Chemistry, BMCUppsala UniversityUppsalaSweden
  2. 2.Department of NeurochemistryStockholm UniversityStockholmSweden

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