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Titlebook: In Vitro Mutagenesis Protocols; Michael K. Trower Book 19961st edition Humana Press 1996

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Modification of the Overlap Extension Method for Extensive Mutagenesis on the Same Template,cient than traditional methods for site-directed mutagenesis (.). However, because synthesis of two complementary primers is required for each mutation, the overlap extension is not the method of choice for extensive mutagenesis of a target sequence.
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Site-Directed Mutagenesis Using Overlap Extension PCR, other complementary regions, which form extensive secondary structures within single-stranded DNA, which often severely decrease the ability to extend annealed primers containing the mutation of interest.
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Site-Directed Mutagenesis Using Double-Stranded Plasmid DNA Templates,cialized vectors, and convenient restriction sites or subcloning of the sequence of DNA to be mutated into a bacteriophage vector like M13 to produce and recover single-stranded DNA. These manipulations present major limitations to the routine use of these methods because they are time consuming and tedious.
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Oligonucleotide-Directed Mutagenesis Using an Improved Phosphorothioate Approach, (.,.) that has been improved to make it quicker and more convenient (.). Selection for the desired mutation takes place in vitro, allowing a high efficiency of mutagenesis to be achieved. Typically, an efficiency of greater than 85% mutants can be obtained with a range of types of mutation.
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Double-Stranded DNA Site-Directed Mutagenesis,and their interactions. Several protocols have been successfully employed to generate such mutants. Here I describe a “linker-scanning” method that I have used to systematically mutate the murine major histocompatibility complex (MHC) class II Eα gene promoter (., ref. .).
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Ligase Chain Reaction for Site-Directed In Vitro Mutagenesis,.). Several techniques have been employed to enhance the frequency of clones containing the desired mutation by selecting against the parental strand through biological means or by in vitro manipulations (.–.).
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In Vitro Mutagenesis Protocols978-1-59259-544-0Series ISSN 1064-3745 Series E-ISSN 1940-6029
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