Abstract
Joining DNA sequences to create linear and circular constructs is a basic requirement in molecular biology. Here we describe PaperClip, a recently developed method, which enables assembly of multiple DNA sequences in one reaction in a combinatorial manner. In contrast to other homology-based multi-part assembly methods currently available, PaperClip allows assembly of a given set of parts in any order without requiring specific single-use oligonucleotides for each assembly order.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Arber W, Linn S (1969) DNA modification and restriction. Annu Rev Biochem 38:467–500. doi:10.1146/annurev.bi.38.070169.002343
Meselson M, Yuan R (1968) DNA restriction enzyme from E. coli. Nature 217(5134):1110–1114
Weiss B, Richardson CC (1967) Enzymatic breakage and joining of deoxyribonucleic acid, I. Repair of single-strand breaks in DNA by an enzyme system from Escherichia coli infected with T4 bacteriophage. Proc Natl Acad Sci U S A 57(4):1021–1028
Engler C, Kandzia R, Marillonnet S (2008) A one pot, one step, precision cloning method with high throughput capability. PLoS One 3(11), e3647. doi:10.1371/journal.pone.0003647
Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S (2011) A modular cloning system for standardized assembly of multigene constructs. PLoS One 6(2), e16765. doi:10.1371/journal.pone.0016765
Shetty R, Lizarazo M, Rettberg R, Knight TF (2011) Assembly of BioBrick standard biological parts using three antibiotic assembly. Methods Enzymol 498:311–326. doi:10.1016/B978-0-12-385120-8.00013-9
Casini A, MacDonald JT, De Jonghe J, Christodoulou G, Freemont PS, Baldwin GS, Ellis T (2014) One-pot DNA construction for synthetic biology: the Modular Overlap-Directed Assembly with Linkers (MODAL) strategy. Nucleic Acids Res 42(1), e7. doi:10.1093/nar/gkt915
Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6(5):343–345. doi:10.1038/nmeth.1318
Quan J, Tian J (2009) Circular polymerase extension cloning of complex gene libraries and pathways. PLoS One 4(7), e6441. doi:10.1371/journal.pone.0006441
Zhang Y, Werling U, Edelmann W (2012) SLiCE: a novel bacterial cell extract-based DNA cloning method. Nucleic Acids Res 40(8), e55. doi:10.1093/nar/gkr1288
Trubitsyna M, Michlewski G, Cai Y, Elfick A, French CE (2014) PaperClip: rapid multi-part DNA assembly from existing libraries. Nucleic Acids Res. doi:10.1093/nar/gku829
Michlewski G, Caceres JF (2010) RNase-assisted RNA chromatography. RNA 16(8):1673–1678. doi:10.1261/rna.2136010
Acknowledgements
This work was supported by Engineering and Physical Sciences Research Council [EP/J02175x/1].
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Trubitsyna, M., Liu, CK., Salinas, A., Elfick, A., French, C.E. (2017). PaperClip: A Simple Method for Flexible Multi-Part DNA Assembly. In: Hughes, R. (eds) Synthetic DNA. Methods in Molecular Biology, vol 1472. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6343-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6343-0_9
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6341-6
Online ISBN: 978-1-4939-6343-0
eBook Packages: Springer Protocols