Optimization of overlap extension PCR for efficient transgene construction
نویسندگان
چکیده
منابع مشابه
Dual Promoter Vector Construction for Simultaneous Gene Expression Using Spliced Overlap Extension by Polymerase Chain Reaction (SOE-PCR) Technique
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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Zinc finger nuclease (ZFN) is a useful tool for endogenous site-directed genome modification. The development of an easier, less expensive and repeatedly usable construction method for various sequences of ZFNs should contribute to the further widespread use of this technology. Here, we establish a novel construction method for ZFNs. Zinc finger (ZF) fragments were synthesized by PCR using shor...
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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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background: causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. brucellosis is the most common zoonotic infectious disease that would cause great economic losses. thus, recognition of pathogenic and immunogenic factors in the genus brucella can lead to control this health problem. objectives: considering the importanc...
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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...
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ژورنال
عنوان ژورنال: MethodsX
سال: 2020
ISSN: 2215-0161
DOI: 10.1016/j.mex.2019.12.001