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QuikChange HT Protein Engineering System Details & Specifications

Overview

 

Site Directed Mutagenesis (SDM) is the method of choice for rationally mutating proteins delivering the highest fidelity and targeted action.

The QuikChange HT Protein Engineering System improves on the proven power of the QuikChange technology by the addition of a high fidelity custom oligonucleotide library. QuikChange HT allows rapid resolution of structural and functional questions by creating libraries of rationally designed mutants, at a price point that enables broad utilization of comprehensive mutant libraries for applications such as single amino acid scanning, site saturations scanning or targeted combinatorial mutagenesis.

 

Save Research Time and Learn More – all in a Single Experiment

Figure 1. QuikChange HT Method

Figure 1. QuikChange HT Method



QC HT Workflow Including Sub-libraries, QuikScan, and QuikCombine

The QuikChange HT Protein Engineering System provides researchers the ability to precisely target and mutate every codon within a 50 amino acid region in less than one day with a single oligo set. Additional oligo sets can mutate several distinct sites e.g. other 50 amino acid regions of the same or of different proteins, in parallel reactions.

This technology provides researchers the ability to design large mutagenic libraries from as few as 1,000 and up to 120,000 user-defined sequences via free access to a mutagenesis workspace software user-interface. The library may consist of numerous oligo sets (up to 20) targeted at distinct regions of a single protein or of multiple proteins. Each oligo set will correspond to a homologous DNA sequence that hybridizes to the same gene fragment and varies in sequence by one or up to four codons. Each DNA sequence contains fully complementary ends for PCR amplification of the oligo set. Each mutagenesis library is customized for each experiment and once received, researchers can generate libraries of transformed competent cells in less than a day.

Figure 3. QuikChange HT Method

Figure 3. QC HT Methods



Validation: Codon Saturation Scanning of GFP

Figure 4. QuikChange HT Method

Figure 4. Codon saturation scanning of GFP. The GFP structure appears to be very sensitive to modification, as seen by the overall low number of mutants that retain fluorescence. The conformation of the beta barrel structure is critical to orientation, maturation, and fluorescence emitted by the chromophore. Other enzymes (e.g., beta gal) appear to be much more tolerant of mutations.

For Research Use Only. Not for use in diagnostic procedures