Randomization of genes by PCR mutagenesis.
نویسندگان
چکیده
منابع مشابه
Randomization of genes by PCR mutagenesis.
A modified polymerase chain reaction (PCR) was developed to introduce random point mutations into cloned genes. The modifications were made to decrease the fidelity of Taq polymerase during DNA synthesis without significantly decreasing the level of amplification achieved in the PCR. The resulting PCR products can be cloned to produce random mutant libraries or transcribed directly if a T7 prom...
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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 QuikChangeTM protocol is one of the simplest and fastest methods for site-directed mutagenesis, but introduces mutations at only one site at a time, and requires two HPLC-purified complementary oligonucleotides. Here, we describe that this method can be used with non-overlapping oligonucleotides. By doing this, two separate sites can be mutagenised simultaneously, or money can be saved by u...
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The polymerase chain reaction (PCR) has been extensively used for the mutation of DNA sequences (1–5,7,8). Many variations of the protocol have been implemented, but most involve a first step in which PCR is used to modify the molecule, and a second step in which the new sequence is subcloned into a vector. A new method, named inverse PCR mutagenesis (IPCRM) has been shown to be an efficient an...
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ژورنال
عنوان ژورنال: Genome Research
سال: 1992
ISSN: 1088-9051
DOI: 10.1101/gr.2.1.28