5989-8097 - 12 Pages

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5989-8097

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Stratagene Mutagenesis Solutions for Your Protein Engineering Needs An Agilent Technologies Company

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Protein engineering via mutagenesis allows researchers to modulate protein activity and characterize structure-function relationships, which enriches our understanding of basic cellular processes and disease mechanisms, fueling discoveries in new therapies for complex diseases such as cancer. Stratagene has been a pioneer in developing innovative mutagenesis tools for the past two decades. Cited in thousands of publications, our QuikChange® site-directed mutagenesis and GeneMorph® random mutagenesis kits are the tools behind many important experiments and scientific...

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Mutagenesis Selection Guide APPLICATION Site-Directed Mutagenesis PRODUCT    DESCRIPTION QuikChange Lightning kit Features faster enzymes and shorter reaction protocols, which provide significant time savings with the same efficiency and accuracy as our original QuikChange kits; suitable for site-directed mutagenesis of all plasmid sizes, and quality-controlled with DNA templates up to 14 kb UNIQUE FEATURES • Protocol completed in <3 hours plus an overnight transformation • Contains the highly processive, ultra-high-fidelity QuikChange Lightning DNA polymerase fusion • Employs QuikSolution...

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Easily Achieve Accurate, Reliable Site-Directed Mutagenesis Site-directed mutagenesis is the method of choice for altering a gene or vector sequence at a selected location. Point mutations, insertions or deletions are introduced by incorporating primers containing the desired modification(s) with a DNA polymerase in an amplification reaction. Stratagene offers the most accurate, efficient and easy-to-use mutagenesis kits available today. Save time with simple, same-day protocol Our QuikChange mutagenesis kits utilize our signature three-step, one-day method to introduce point mutations,...

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Optimized for your success QuikChange kits include all required reagents, including controls, so you have everything you need. Enzymes, buffers and dNTPs are optimized, saving you time and ensuring consistent results. In addition, all of our kits include Dpn I enzyme and high-efficiency competent cells. Primer design made easy The QuikChange Primer Design Program assists you in designing mutagenic primers for your QuikChange site-directed mutagenesis experiments. • Generates appropriate primer sequences for point mutations, deletions and insertions, with optimal melting temperature and free...

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Stratagene recognizes the important role of site-directed mutagenesis in your research. That is why we continue to develop new, more powerful tools to help you perform your experiments more easily and in less time, in order to address your most challenging applications. QuikChange Lightning site-directed mutagenesis kit: a powerful new tool Introduce point mutations, insertions or deletions in less than three hours followed by an overnight transformation, while maintaining the high accuracy and efficiency that you've come to expect with QuikChange kits (Figure 2). Using our signature...

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QuikChange II site-directed mutagenesis kit: proven technology Achieve efficient and accurate site-directed mutagenesis with our QuikChange II kits, featuring PfuUltra DNA polymerased, a genetically engineered mutant of Pfu DNA polymerase with enhanced proofreading capability. QuikChange II XL: For your most challenging applications Our QuikChange II XL site-directed mutagenesis kita was created for efficient mutagenesis of large or otherwise difficult to mutagenize plasmid templates. The QuikChange II XL kit features components specifically designed for more efficient DNA replication and...

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Number of sites mutated Efforts to elucidate protein structure-function relationships typically begin by identifying key residues through predictions from structural data, identifying changes in activity accompanying single-site mutagenesis9 or identifying sequence changes in mutants with altered activity isolated from random mutant libraries.10 Once you identify key residues, you may need to create numerous multi-site mutants to determine the effects of combining key mutations. Our QuikChange Multi site-directed mutagenesis kit provides a simple method for introducing mutations at up to...

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As shown in Figure 5B, a key phosphorylation site (tyrosine) was subject to saturation mutagenesis by incorporating a degenerate-(NNN)-codon primer with the QuikChange Multi kit. This example shows three (W, Q, R) of the 19 mutants that are produced in a single reaction. As above, the resulting mutant clone collection can be screened as a library to identify changes in activity, or as individual clones to precisely characterize structure-function relationships. Additional studies have shown that the number of clones per QuikChange Multi reaction is sufficient for creating fully...

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Random mutagenesis is often the first step on the path to creating a preliminary map of your protein's functional domains. Typically, a gene or gene fragment is randomly mutated by error-prone PCR, mutant sequences are cloned into a suitable expression vector, and libraries are screened to identify clones that exhibit changes in protein function. By correlating changes in activity to changes in amino acid sequence, researchers can begin to identify functional domains and map structure-function relationships. Our GeneMorph products facilitate the process of random mutagenesis by providing...

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As shown in Figure 7, domains are first amplified to introduce random mutations using error-prone PCR with Mutazyme II DNA polymerase. The purified PCR products then serve as megaprimers for the EZClone reaction. They are denatured and annealed to the original donor plasmid and then extended with EZClone high-fidelity DNA polymerase. This reaction is thermal cycled several times and treated with Dpn I enzyme to remove background DNA before transformation into competent E. coli. After transformation, the resulting mutant library can be screened in your functional assay. Mutant Megaprimer...

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