title: De Novo Assembly of Plasmids Using Yeast Recombinational Cloning.
abstract: Molecular cloning is a cornerstone of modern biology laboratories. However, traditional cloning can be time-consuming and problematic. We outline herein a method that utilizes the endogenous gap repair system of yeast cells to clone and assemble DNA constructs. This system is simple, cheap, and requires minimal reagents. It can be used for the assembly of both simple (single DNA fragments) and complex (multiple DNA fragments) constructs into plasmids.
authors.affiliation: Department of Biochemistry and Microbiology, Rutgers University, 76 Lipman Dr., New Brunswick, NJ, 08904, USA.
authors.forename: Ameya A
authors.initials: AA
authors.lastname: Mashruwala
authors.affiliation: Department of Biochemistry and Microbiology, Rutgers University, 76 Lipman Dr., New Brunswick, NJ, 08904, USA. jmboyd@AESOP.Rutgers.edu.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Boyd
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D059765:Homologous Recombination / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013210:Staphylococcus / Q000235:genetics* / Q000472:pathogenicity
keywords: Genetics; Homologous recombination; Molecular biology; Molecular cloning; Staphylococcus; Yeast