title: Ex vivoDNA Assembly.
abstract: 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 powerfulin vitroandin vivoassembly of DNA. However, monetary and time costs have limited these approaches. Here, we report anex vivoDNA assembly method that uses cellular lysates derived from a commonly used laboratory strain ofEscherichia colifor 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.
authors.affiliation: Integrative Life Sciences Program, Virginia Commonwealth University , Richmond, VA , USA.
authors.forename: Adam B
authors.initials: AB
authors.lastname: Fisher
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, VA , USA.
authors.forename: Zachary B
authors.initials: ZB
authors.lastname: Canfield
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, VA , USA.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Hayward
authors.affiliation: Integrative Life Sciences Program, Virginia Commonwealth University , Richmond, VA , USA ; Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, VA , USA.
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Fong
authors.affiliation: Department of Chemical and Life Science Engineering, Virginia Commonwealth University , Richmond, VA , USA.
authors.forename: George H
authors.initials: GH
authors.lastname: McArthur
keywords: DNA assembly; cellular lysates; colorimetric screen; end joining; ex vivo; genetic engineering; synthetic biology