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: Hasnain, Hussain, Chong, Nikson Fatt Ming
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2016
Subjects:
Online Access:http://ir.unimas.my/id/eprint/14294/
http://ir.unimas.my/id/eprint/14294/1/Combined%20Overlap%20Extension%20PCR%20Method%20for%20Improved%20Site%20Directed%20Mutagenesis%20%28abstract%29.pdf
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author Hasnain, Hussain
Chong, Nikson Fatt Ming
author_facet Hasnain, Hussain
Chong, Nikson Fatt Ming
author_sort Hasnain, Hussain
building UNIMAS Institutional Repository
collection Online Access
description 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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institution Universiti Malaysia Sarawak
institution_category Local University
language English
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publishDate 2016
publisher Hindawi Publishing Corporation
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spelling unimas-142942017-02-17T02:43:35Z http://ir.unimas.my/id/eprint/14294/ Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis Hasnain, Hussain Chong, Nikson Fatt Ming Q Science (General) QR Microbiology 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. Hindawi Publishing Corporation 2016 Article PeerReviewed text en http://ir.unimas.my/id/eprint/14294/1/Combined%20Overlap%20Extension%20PCR%20Method%20for%20Improved%20Site%20Directed%20Mutagenesis%20%28abstract%29.pdf 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 https://www.hindawi.com/journals/bmri/
spellingShingle Q Science (General)
QR Microbiology
Hasnain, Hussain
Chong, Nikson Fatt Ming
Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title_full Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title_fullStr Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title_full_unstemmed Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title_short Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
title_sort combined overlap extension pcr method for improved site directed mutagenesis
topic Q Science (General)
QR Microbiology
url http://ir.unimas.my/id/eprint/14294/
http://ir.unimas.my/id/eprint/14294/
http://ir.unimas.my/id/eprint/14294/1/Combined%20Overlap%20Extension%20PCR%20Method%20for%20Improved%20Site%20Directed%20Mutagenesis%20%28abstract%29.pdf