Random Mutagenesis by Whole-Plasmid PCR Amplification
نویسندگان
چکیده
منابع مشابه
Random mutagenesis by whole-plasmid PCR amplification.
Random mutagenesis has become a powerful means of studying the effects of a large number of permutations of a particular DNA sequence. As a prime example, libraries of randomized cDNA clones, when translated into their corresponding proteins, can be useful in investigating the functional contributions of a mutagenized region to the overall properties of a protein. Existing molecular cloning tec...
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The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it. Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector need optimizing. Even then, the resultant library is often plagued with unwanted plasmids that have no inserts or multiple inserts. These species have to be eradicated to...
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INTRODUCTIONPCR-based whole-genome amplification (WGA) has the goal of generating microgram quantities of genome-representative DNA from picogram or nanogram amounts of starting material. This amplification should introduce little, or ideally no, representational bias. Unlike other techniques for WGA, PCR-based methods are generally less affected by DNA quality and are more applicable to DNA ex...
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In vitro random mutagenesis is a powerful tool for altering properties of enzymes. We describe here a novel random mutagenesis method using rolling circle amplification, named error-prone RCA. This method consists of only one DNA amplification step followed by transformation of the host strain, without treatment with any restriction enzymes or DNA ligases, and results in a randomly mutated plas...
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We thank John Connolly for helpful discussions and critical reading of the manuscript. We are very grateful to Fabrice Chareyre, Laetitia Gressin, Catherine Giudicelli, Jean Christophe Beaudoin and Isabelle Legall for help in the BAC screening, sequencing and genotyping. This work was supported by a grant from the Association pour la Recherche sur le Cancer (ARC) and the Ministère de l’Educatio...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1998
ISSN: 0736-6205,1940-9818
DOI: 10.2144/98243st01