Efficient random mutagenesis method with adjustable mutation frequency by use of PCR and dITP
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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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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We have developed an efficient method for site-directed mutagenesis using two subsequential rounds of PCR. In this method, PCR conditions are optimized to favor high fidelity of Taq DNA polymerase in the presence of equimolar concentrations of MgCI2 and dNTP in the reaction mixture (pH 5.5-6.2). This method makes use of a pair of universal primers and the multiple cloning site of pUC/M13 vector...
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Procedures to introduce point mutations into specific DNA fragments are important tools to study gene function (2,10). UV light and chemical mutagens have been used in vivo to increase the frequency of random mutagenesis for specific DNA targets carried on plasmid or phage vectors being propagated in growing cells (2,11). Both methods induced mutations in the target DNA as well as in vector and...
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هدف از این تحقیق بررسی برخی عوامل ادراکی واحساسی یعنی استفاده از شیوه های یادگیری زبان ، انگیزه ها ونگرش نسبت به زبان انگلیسی در رابطه با زمینه زبانی زبان آموزان می باشد. هدف بررسی این نکته بود که آیا اختلافی چشمگیر میان زبان آموزان دو زبانه و تک زبانه در میزان استفاده از شیوه های یادگیری زبان ، انگیزه ها نگرش و سطح مهارت زبانی وجود دارد. همچنین سعی شد تا بهترین و موثرترین عوامل پیش بینی کننده ...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1993
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/21.3.777