AUTHOR=Fisher Adam B., Canfield Zachary B., Hayward Laura C., Fong Stephen , McArthur George H.

TITLE=Ex vivo DNA Assembly

JOURNAL=Frontiers in Bioengineering and Biotechnology

VOLUME=1

YEAR=2013

URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2013.00012

DOI=10.3389/fbioe.2013.00012

ISSN=2296-4185

ABSTRACT=<p>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 <italic>in vitro</italic> and <italic>in vivo</italic> assembly of DNA. However, monetary and time costs have limited these approaches. Here, we report an <italic>ex vivo</italic> DNA assembly method that uses cellular lysates derived from a commonly used laboratory strain of <italic>Escherichia coli</italic> 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.</p>