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DNA Assembly and Cloning

For the purposes of cloning, DNA assembly refers to a method to physically link together multiple fragments of DNA, in an end-to-end fashion, to achieve a desired, higher-order assembly prior to joining to a vector. This process is the cornerstone of the synthetic biology movement, and allows the construction of novel biological systems and devices using defined components. The methods are carried out in vitro, and are typically enzymatically driven with the final constructions being maintained in microbial host cells.

To help select the best DNA assembly method for your needs, please use our Synthetic Biology/DNA Assembly Selection Chart.

Learn more about the various DNA Assembly methods: NEBuilder HiFi DNA Assembly, Gibson AssemblyBioBrick AssemblyGolden Gate Assembly


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DNA Assembly and Cloning includes these areas of focus:
NEBuilder® HiFi DNA Assembly
Gibson Assembly®
BioBrick® Assembly
Golden Gate Assembly
Protocols for DNA Assembly and Cloning
Application Notes for DNA Assembly and Cloning
    Publications related to DNA Assembly and Cloning
    • Anton, B.P., Morgan, R.D., Ezraty, B., Manta, B., Barras, F., Berkmen, M. (2019) Complete genome sequence of Escherichia coli BE104, an MC4100 drivative lacking the methionine reductive pathway Microbiol Resour Announc; 8 (29), e00721-19. PubMedID: 31296691, DOI: 10.1128/MRA.00721-19
    • Potapov, V., Ong, J.L., Kucera, R.B., Langhorst, B.W., Bilotti, K., Pryor, J.M., Cantor, E.J., Canton, B., Knight, T.F., Evans, T.C., Lohman, G.J.S. (2018) Comprehensive profiling of four base overhang ligation fidelity by T4 DNA ligase and application to DNA assembly ACS Synth Biol; 7 (11), PubMedID: 30335370, DOI: 10.1021/acssynbio.8b00333
    • Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske M (2014) A Modular Plasmid Assembly Kit for Multigene Expression, Gene Silencing and Silencing Rescue in Plants PLoS One; 9(2), e88218. PubMedID: 24551083, DOI: 10.1371/journal.pone.0088218
    • Abil Z, Denard CA, Zhao H (2014) Modular assembly of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA J Biol Eng; 8(1), 7. PubMedID: 24581042, DOI: 10.1186/1754-1611-8-7
    • Feng Y, Zhang S, Huang X (2014) A robust TALENs system for highly efficient mammalian genome editing Sci Rep; 4, 3632. PubMedID: 24407151, DOI: 10.1038/srep03632
Legal Information

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.


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