title: Evaluation of seamless ligation cloning extract preparation methods from an Escherichia coli laboratory strain.
abstract: Seamless ligation cloning extract (SLiCE) is a simple and efficient method for DNA cloning without the use of restriction enzymes. Instead, SLiCE uses homologous recombination activities from Escherichia coli cell lysates. To date, SLiCE preparation has been performed using an expensive commercially available lytic reagent. To expand the utility of the SLiCE method, we evaluated different methods for SLiCE preparation that avoid using this reagent. Consequently, cell extracts prepared with buffers containing Triton X-100, which is a common and low-cost nonionic detergent, exhibited sufficient cloning activity for seamless gene incorporation into a vector.
authors.affiliation: Department of Bioresource and Environmental Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kita-ku, Kyoto 603-8555, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Okegawa
authors.affiliation: Department of Bioresource and Environmental Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kita-ku, Kyoto 603-8555, Japan. Electronic address: motohas@cc.kyoto-su.ac.jp.
authors.forename: Ken
authors.initials: K
authors.lastname: Motohashi
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial
keywords: Detergent extraction; Homologous recombination; SLiCE; Seamless DNA cloning