The original QuikChange Site-Directed Mutagenesis Kits speed up and simplify site-directed mutagenesis studies. The kits eliminate the need for sub-cloning into M13-based bacteriophage vectors and for ss-DNA rescue. This allows oligo-mediated introduction of site-specific mutations into virtually any double-stranded plasmid DNA. In addition, the XL version of the kit is specially designed for efficient mutagenesis of large (4 -14 kb) or otherwise difficult-to mutagenize plasmid templates. The XL kit features components specifically designed for more efficient DNA replication and bacterial transformation.
- >80% mutagenesis efficiency for one site; >55% mutagenesis efficiency for three mutations simultaneously
- Efficient and accurate linear amplification minimizes unwanted errors
QuikChange Site-Directed Mutagenesis Kit: This kit uses PfuTurbo DNA polymerase. It replicates plasmid strands with high fidelityll without displacing mutant oligonucleotide primers. The process uses a supercoiled dsDNA vector with insert and two synthetic oligo primers containing the mutation. Incorporating oligo primers creates a mutated plasmid with staggered nicks. The product is treated with Dpn I after temperature cycling. Dpn I digests the parental DNA template, selecting mutation-containing synthesized DNA. The nicked vector DNA containing the mutations is transformed into XL1-Blue supercompetent cells.
QuikChange XL Site-Directed Mutagenesis Kit: The QuikChange II XL site-directed mutagenesis kit specializes in efficient mutagenesis of large (8–14 kb) or otherwise difficult-to mutagenize plasmid templates and features components designed for more efficient DNA replication and bacterial transformation. A QuikSolution reagent is provided to facilitate replication of large plasmids, while XL10-Gold Ultracompetent Cells have been included to ensure the highest transformation efficiencies possible. XL10-Gold cells contain the Hte phenotype, increasing the transformation efficiency of larger DNA plasmids.
QuikChange Multi site-directed mutagenesis kit: The QuikChange Multi site-directed mutagenesis system is a novel technology that allows mutagenesis at multiple sites in a single round, using a single oligonucleotide per site. This system simplifies randomizing key amino acids using oligos containing degenerate codons. No specialized vectors or unique restriction sites are needed to use the kit; virtually any plasmid up to 8 kb is a suitable template. The rapid three-step procedure introduces mutations at three different sites simultaneously in the 4 kb QuikChange Multi control plasmid with >50% efficiency.
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