Unit

UNIT 8.2B Gene Synthesis: Assembly of Target Sequences Using Mutually Priming Long Oligonucleotides

  1. David D. Moore

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0802bs26

Current Protocols in Molecular Biology

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.

Author Information

  1. Massachusetts General Hospital, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 1994

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.