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Improved GPCR Engineering With Synthetic Site Saturation Variant Libraries

Protein engineering and directed evolution applications often use saturation mutagenesis to create a library of variants for screening. However, saturation mutagenesis by error-prone PCR (epPCR) suffers from amplification biases and incomplete access to the codon mutational space. Site Saturation Variant Libraries (SSVLs) offer a synthetic alternative free of the technical limitations of epPCR. Here, the performance of a Twist SSVL was benchmarked against an epPCR library using a glucose activation assay in yeast. Compared to epPCR, the SSVL produced greater variant representation while also simplifying downstream validation steps by providing access to the complete variant diversity.

Download this app note to learn more about:

  • How saturation mutagenesis can be used to identify functional GPCR variants
  • Why error-prone PCR results in low variant representation
  • The advantages of synthetic saturation mutagenesis libraries