title: PaperClip: rapid multi-part DNA assembly from existing libraries.
abstract: Assembly of DNA 'parts' to create larger constructs is an essential enabling technique for bioengineering and synthetic biology. Here we describe a simple method, PaperClip, which allows flexible assembly of multiple DNA parts from currently existing libraries cloned in any vector. No restriction enzymes, mutagenesis of internal restriction sites, or reamplification to add end homology are required. Order of assembly is directed by double stranded oligonucleotides-'Clips'. Clips are formed by ligation of pairs of oligonucleotides corresponding to the ends of each part. PaperClip assembly can be performed by polymerase chain reaction or by cell extract-mediated recombination. Once multi-use Clips have been prepared, assembly of at least six DNA parts in any order can be accomplished with high efficiency within several hours.
authors.affiliation: School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK School of Engineering, University of Edinburgh, Edinburgh, EH9 3JL, UK M.Trubitsyna@ed.ac.uk.
authors.forename: Maryia
authors.initials: M
authors.lastname: Trubitsyna
authors.affiliation: School of Biological Sciences, Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3JR, UK.
authors.forename: Gracjan
authors.initials: G
authors.lastname: Michlewski
authors.affiliation: School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK.
authors.forename: Yizhi
authors.initials: Y
authors.lastname: Cai
authors.affiliation: School of Engineering, University of Edinburgh, Edinburgh, EH9 3JL, UK.
authors.forename: Alistair
authors.initials: A
authors.lastname: Elfick
authors.affiliation: School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK M.Trubitsyna@ed.ac.uk.
authors.forename: Christopher E
authors.initials: CE
authors.lastname: French
mesh_terms: D004247:DNA / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D009841:Oligonucleotides; D004247:DNA