UNIT 8.2B Gene Synthesis: Assembly of Target Sequences Using Mutually Priming Long Oligonucleotides
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Moore, D. D. 2001. Gene Synthesis: Assembly of Target Sequences Using Mutually Priming Long Oligonucleotides. Current Protocols in Molecular Biology. 26:8.2B:8.2.8–8.2.13.
- Published Online: 1 MAY 2001
- Published Print: APR 1994
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.