title: Seamless Ligation Cloning Extract (SLiCE) cloning method.
abstract: SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies.
authors.affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY, USA.
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Zhang
authors.forename: Uwe
authors.initials: U
authors.lastname: Werling
authors.forename: Winfried
authors.initials: W
authors.lastname: Edelmann
mesh_terms: D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics