Details

Autor(en) / Beteiligte

• Hussain, Mohd Hasnain Md
• Chong, Nikson Fatt-Ming
• Ongecha, John M.

Titel

Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis

Ist Teil von

• BioMed research international, 2016-01, Vol.2016 (2016), p.1-7

Ort / Verlag

Cairo, Egypt: Hindawi Publishing Corporation

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.