Gene synthesis: assembly of target sequences using mutually priming long oligonucleotides
1Massachusetts General Hospital, Boston, Massachusetts, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Abstract:
This protocol uses pairs of oligonucleotides annealed at a short duplex segment at their 3' ends as both templates and primers (mutually primed synthesis) to generate desired sequences up to 400 bp in a single step. The procedure is divided into three sections: design of the oligonucleotides, extension by mutually primed synthesis, and cloning of the extension products. The strategy for design of oligonucleotides is discussed in the commentary.
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