Precise large deletions by the PCR-based overlap extension method
نویسندگان
چکیده
منابع مشابه
A Nested-Splicing by Overlap Extension PCR Improves Specificity of this Standard Method
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...
متن کاملCombined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
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 nested-splicing by overlap extension pcr improves specificity of this standard method
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 thermoca...
متن کاملRepeatable Construction Method for Engineered Zinc Finger Nuclease Based on Overlap Extension PCR and TA-Cloning
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...
متن کاملA Nested-Splicing by Overlap Extension PCR Improves Specificity of this Standard Method.
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 thermoc...
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ژورنال
عنوان ژورنال: Molecular Biotechnology
سال: 1995
ISSN: 1073-6085,1559-0305
DOI: 10.1007/bf02907467