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

Abstract

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.

Inventors:
 [1];  [2];  [3];  [4];  [5]
  1. San Leandro, CA
  2. (Danville, CA)
  3. Tracy, CA
  4. Berkeley, CA
  5. Livermore, CA
Issue Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1013053
Patent Number(s):
7,871,799
Application Number:
10/727,779
Assignee:
Lawrence Livermore National Security, LLC (Livermore, CA) OAK
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Gardner, Shea N, Mariella, Jr., Raymond P., Christian, Allen T, Young, Jennifer A, and Clague, David S. Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions. United States: N. p., 2011. Web.
Gardner, Shea N, Mariella, Jr., Raymond P., Christian, Allen T, Young, Jennifer A, & Clague, David S. Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions. United States.
Gardner, Shea N, Mariella, Jr., Raymond P., Christian, Allen T, Young, Jennifer A, and Clague, David S. Tue . "Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions". United States. https://www.osti.gov/servlets/purl/1013053.
@article{osti_1013053,
title = {Sequential addition of short DNA oligos in DNA-polymerase-based synthesis reactions},
author = {Gardner, Shea N and Mariella, Jr., Raymond P. and Christian, Allen T and Young, Jennifer A and Clague, David S},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2011},
month = {1}
}

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