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QuikChange Lightning Multi Site-Directed Mutagenesis Kit 多点突变试剂盒 210515 推荐

更多点突变试剂盒

  • Greater than 55% mutagenesis efficiency for 3 mutations simultaneously, capable of five mutations simultaneously
  • Easy three step non-PCR procedure is now even faster – complete your multi-site mutagenesis experiment in 3 hours.
  • Uses highly processive ultra-high-fidelity QuikChange Lightning DNA polymerase fusion enzyme
  • Click here to use our free, convenient QuikChange® Primer Design Program today.
  • Download the Science Mutagenesis Techniques Collections sponsored by the Stratagene Products Division of Agilent Technologies.

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Multiple Site Mutations with New Enzyme Blend

The QuikChange Lightning Multi Site-Directed Mutagenesis Kit offers the most rapid and reliable method for site-directed mutagenesis of plasmid DNA at up to five different sites simultaneously. Using the most advanced high fidelity enzyme technology, the protocols have been accelerated while maintaining the highest accuracy for site-directed mutagenesis. The QuikChange Lightning Multi Site-Directed Mutagenesis Kit delivers results up to three times faster than our original multi-site mutagenesis kit.
 

As with our original QuikChange Multi kit, a single mutagenic oligonucleotide is required to mutagenize each site, using a double-stranded DNA template. The kit’s exclusive enzyme blend features a unique Pfu Fusion-based DNA polymerase,** providing high fidelity DNA synthesis. The rapid PCR cycling parameters of the unique enzyme blend, together with our newly optimized Dpn I enzyme, allow for multi-site mutagenesis inapproximately 3 hours.

Ultra Rapid Protocol

The QuikChange Lightning Multi Site-Directed technology provides significant time savings during linear amplification and digestion of the parental DNA target. The additional time savings allows researchers to easily complete the protocol in a single day and transform the newly mutated DNA construct overnight. In the case of a 5kb target template QuikChange Lightning Multi is 4x faster than the original QuikChange Multi kit.

Single-Day Multi-Site Mutagenesis Saves Time

The QuikChange Lightning Multi kit provides a quick and efficient method for carrying out site-directed mutagenesis in up to 5 sites simultaneously. Incorporating multiple mutations at up to 5 sites in a single reaction results in nearly two weeks of time and effort saved compared to sequential mutagenesis methods.

Accelerate Discovery Using Saturation Mutagenesis

Saturation mutagenesis provides a much more comprehensive analysis of structure-function relationships compared to single amino acid replacements or error-prone PCR. Strategies using traditional error-prone PCR typically use stressful PCR conditions to randomly generate single-base changes throughout a gene sequence. However, a large number of mutations types are not represented adequately in random mutant collections, in particular, mutations requiring 2-3 base pair changes per codon. To access a larger fraction of protein sequence space, site-specific saturation mutagenesis is commonly used to introduce all possible mutations at key sites or adjacent sites. Using the QuikChange Lightning Multi kit for saturation mutagenesis is an efficient and effective method to change substrate specificity, thermostability or solvent resistance, making it fast and efficient compared to other techniques.

Engineered Mutant Clone Collections

Once you identify key residues through whole-gene random mutagenesis and screening, you may need to create numerous multi-site mutants to determine the effects of combining key mutations. This task can be daunting with respect to both time and reagent cost, depending upon the number of sites you are investigating. In a single QuikChange Multi reaction with multiple primers, you can create a complete, diverse set of mutants called an Engineered Mutant Clone collection.

Elegant Non-PCR Method Delivers High Accuracy and Efficiency

The QuikChange Multi kit technology employs a non-PCR method, which drastically reduces unwanted second-site mutations. Coupling this method with our high-fidelity DNA polymerase minimizes unwanted errors during mutant strand synthesis while delivering high mutagenesis efficiency.

Easy to Use, 3-Step Procedure

Primers containing mutations are synthesized to only one strand of the double-stranded template. Our unique enzyme extends the mutagenic primers with the highest possible accuracy under non-strand displacing conditions, generating one strand bearing multiple mutations and nicks. The nicks are then sealed by the unique QuikChange Multi enzyme blend. This reaction is treated with Dpn I to digest the parental DNA template; thus, enriching for the multiply mutated single-stranded DNA. The mixture is then transformed into XL10-Gold ultracompetent cells, where the mutant single-stranded DNA is converted into duplex form in vivo. The mutagenic efficiency is approximately 50% using the control system with three mutagenic primers and up to 95% efficient using a single mutagenic oligo.

Multi Site-Directed Mutagenesis Applications

  • Determine structure-function relationships

  • Alter enzyme substrate specificity or kinetics

  • Define protein-protein interaction domains

  • Characterize promoter or DNA element structure

  • Improve protein antigenicity, activity and stability

  • Facilitate x-ray crystallography studies

 


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QuikChange Lightning Mutagenesis Kit

质粒<14kb

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QuikChange XL Site-Directed Mutagenesis Kit

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QuikChange II XL Site-Directed Mutagenesis Kit

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12591

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随机

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GeneMorph II Random Mutagenesis Kit 随机突变克隆 200550 10T 8310

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Dpn I Restriction E500402.pdfnzyme

配套试剂

500402

200U

1848

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    体外定点突变技术是研究蛋白质结?#36141;?#21151;能之间复杂关系的有力工具,也是实验室中改造和优化基因常用的手段,对某个已知基因的特定碱基进行定点改变、缺失或者插入可以改变对应氨基酸序列和蛋白质结构,可以进一步了解蛋白质结?#36141;?#21151;能之间的关系,探讨蛋白质的结构域。上海前尘公司常备著名品牌Stratagene的各种定点突变检测试剂盒,并提供专业的技术支持,使您科研无忧!

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