Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis

Hasnain, Hussain and Chong, Nikson Fatt Ming (2016) Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis. BioMed Research International, 2016. ISSN 2314-6133

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Abstract

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.

Item Type: Article
Uncontrolled Keywords: combined overlap extension PCR method, Site Directed Mutagenesis technique, Mutagenesis, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, research, Universiti Malaysia Sarawak
Subjects: Q Science > Q Science (General)
Q Science > QR Microbiology
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 16 Nov 2016 02:24
Last Modified: 17 Feb 2017 02:43
URI: http://ir.unimas.my/id/eprint/14294

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