Combined 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 re...
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| Main Authors: | , |
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| Format: | E-Article |
| Language: | English |
| Published: |
Hindawi Publishing Corporation
2016
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/14294/1/Combined%20Overlap%20Extension%20PCR%20Method%20for%20Improved%20Site%20Directed%20Mutagenesis%20%28abstract%29.pdf http://ir.unimas.my/id/eprint/14294/ https://www.hindawi.com/journals/bmri/ |
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| Summary: | 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 carried out, with each experiment mutating between two to six base pairs. Up to four adjacent codons were changed in a single experiment. This method is especially useful for codon optimization, where doublet or triplet rare codons can be changed using a single mutagenic primer-set, in a single experiment. |
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