Even with decreasing DNA synthesis costs there remains a need for inexpensive, rapid, and reliable methods for assembling synthetic DNA into larger constructs or combinatorial libraries. Advances in cloning techniques have resulted in powerful in vitro and in vivo assembly of DNA. However, monetary and time costs have limited these approaches. Here, we report an ex vivo DNA assembly method that uses cellular lysates derived from a commonly used laboratory strain of Escherichia coli for joining double-stranded DNA with short end homologies embedded within inexpensive primers. This method concurrently shortens the time and decreases costs associated with current DNA assembly methods.
Keywords: DNA assembly, ex vivo, end joining, cellular lysates, colorimetric screen, synthetic biology, genetic engineering
Citation: Fisher AB, Canfield ZB, Hayward LC, Fong SS and McArthur GH IV (2013) Ex vivo DNA assembly. Front. Bioeng. Biotechnol. 1:12. doi: 10.3389/fbioe.2013.00012
Received: 06 September 2013; Accepted: 06 October 2013;
Published online: 21 October 2013.
Edited by:Zhanglin Lin, Tsinghua University, China
Reviewed by:Weiwen Zhang, Tianjin University, China
Copyright: © 2013 Fisher, Canfield, Hayward, Fong and McArthur IV. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Stephen S. Fong and George H. McArthur IV, Department of Chemical and Life Science Engineering, Virginia Commonwealth University, 601 West Main Street, Richmond, VA 23284-3028, USA e-mail: firstname.lastname@example.org; email@example.com