Chimerization of multiple antibody classes using splice overlap extension PCR
نویسندگان
چکیده
منابع مشابه
Simultaneous introduction of multiple mutations using overlap extension PCR.
Introduction of multiple mutations can be accomplished through phage M13-based site-directed mutagenesis using several oligonucleotides (6). The major drawback of this method is the low efficiency of generating all the intended mutations simultaneously (7,9). The alternative methods for introducing multisite mutations are based on polymerase chain reaction (PCR) (1) and ligase chain reaction (L...
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There are two different co-expression systems including bicistronic; dual-vector or two-promoter to express two different genes simultaneously and also to study protein-protein interactions. Bicistronic system has disadvantages e.g. compared with two-promoter system. In this paper, a simple method based on spliced overlap extension by polymerase chain reaction (SOE-PCR) technique was demonstrat...
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Background: Splicing by overlap extension (SOE) PCR is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function. Objectives: We introduced a nested-SOE-PCR (N –SOE-PCR) in order to increase the specificity and generating mutations in a gene by SOE-PCR. Materials and Methods: Genomic DNA from Bacillus thermo...
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The combined overlap extension PCR (COE-PCR) method developed in this work combines the strengths of the overlap extension PCR (OE-PCR) method with the speed and ease of the asymmetrical overlap extension (AOE-PCR) method. This combined method allows up to 6 base pairs to be mutated at a time and requires a total of 40-45 PCR cycles. A total of eight mutagenesis experiments were successfully ca...
متن کاملA rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.
A rapid method is described to efficiently perform site-directed mutagenesis based on overlap extension polymerase chain reaction (OE-PCR). Two template DNA molecules in different orientations relative to only one universal primer were amplified in parallel. By choosing a high dilution of mutagenic primers it was possible to run an overlap extension PCR in only one reaction without purification...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 2005
ISSN: 0736-6205,1940-9818
DOI: 10.2144/05382bm01