An improved overlap extension PCR for simultaneous multiple sites large fragments insertion, deletion and substitution

Simultaneous introduction of multiple mutations using overlap extension PCR.

Introduction of multiple mutations can be accomplished through phage M13-based site-directed mutagenesis using several oligonucleotides (6). The major drawback of this method is the low efficiency of generating all the intended mutations simultaneously (7,9). The alternative methods for introducing multisite mutations are based on polymerase chain reaction (PCR) (1) and ligase chain reaction (L...

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 requires a total of 40-45 PCR cycles. A total of eight mutagenesis experiments were successfully ca...

Dual Promoter Vector Construction for Simultaneous Gene Expression Using Spliced Overlap Extension by Polymerase Chain Reaction (SOE-PCR) Technique

There are two different co-expression systems including bicistronic; dual-vector or two-promoter to express two different genes simultaneously and also to study protein-protein interactions. Bicistronic system has disadvantages e.g. compared with two-promoter system. In this paper, a simple method based on spliced overlap extension by polymerase chain reaction (SOE-PCR) technique was demonstrat...

Marker Codes for Channels with Insertion, Deletion and Substitution Errors

...................................................................................................................... 3

Versatile PCR-mediated insertion or deletion mutagenesis.