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Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions

United States Patent

January 18, 2011
View the Complete Patent at the US Patent & Trademark Office
Lawrence Livermore National Laboratory - Visit the Industrial Partnerships Office Website
A method of fabricating a DNA molecule of user-defined sequence. The method comprises the steps of preselecting a multiplicity of DNA sequence segments that will comprise the DNA molecule of user-defined sequence, separating the DNA sequence segments temporally, and combining the multiplicity of DNA sequence segments with at least one polymerase enzyme wherein the multiplicity of DNA sequence segments join to produce the DNA molecule of user-defined sequence. Sequence segments may be of length n, where n is an even or odd integer. In one embodiment the length of desired hybridizing overlap is specified by the user and the sequences and the protocol for combining them are guided by computational (bioinformatics) predictions. In one embodiment sequence segments are combined from multiple reading frames to span the same region of a sequence, so that multiple desired hybridizations may occur with different overlap lengths. In one embodiment starting sequence fragments are of different lengths, n, n+1, n+2, etc.
Gardner; Shea N. (San Leandro, CA), Mariella, Jr.; Raymond P. (Danville, CA), Christian; Allen T. (Tracy, CA), Young; Jennifer A. (Berkeley, CA), Clague; David S. (Livermore, CA)
Lawrence Livermore National Security, LLC (Livermore, CA)
10/ 727,779
December 3, 2003
The United States Government has rights in this invention pursuant to Contract No. W-7405-ENG-48 between the United States Department of Energy and the University of California for the operation of Lawrence Livermore National Laboratory.