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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 Assembly, BioBrick Assembly, Golden 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
  1. 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), PubMedID: 31296691, DOI: 10.1128/MRA.00721-19
  2. 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 Synthetic Biology. 7 (11), PubMedID: 30335370, DOI: 10.1021/acssynbio.8b00333
  3. 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), PubMedID: 24551083, DOI: 10.1371/journal.pone.0088218
  4. Feng Y, Zhang S, Huang X 2014. A robust TALENs system for highly efficient mammalian genome editing Sci Rep. 4, PubMedID: 24407151, DOI: 10.1038/srep03632
  5. 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), PubMedID: 24581042, DOI: 10.1186/1754-1611-8-7
Legal Information

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@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.


Videos

  1. NEBTVwebinar_NEBuilderHiFi_720

    NEBuilder HiFi: the next generation of DNA Assembly

    In this webinar, we will explore some of the latest applications for NEBuilder HiFi DNA Assembly. View full Q & A summary.

  2. Construction

    Construction of an sgRNA-Cas9 Expression Vector via an ssOligo Bridge

    Learn how you can use NEBuilder HiFi to generate an sgRNA-Cas9 expression vector with a single-stranded oligo bridge.

  3. nebuilder_overview_thumb

    Introduction to NEBuilder HiFi DNA Assembly

    Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB Gibson Assembly®.

  4. NEBTVwebinar_NEBuilderHiFi_720

    NEBuilder HiFi: the next generation of DNA Assembly

    In this webinar, we will explore some of the latest applications for NEBuilder HiFi DNA Assembly. View full Q & A summary.

  5. nebuilder_3primemismatchvideo_thumb

    NEBuilder® HiFi DNA Assembly Removal of 3´ end Mismatches

    Did you know that one of the advantages of NEBuilder HiFi is that it removes 3’ and 5’ -end mismatch sequences prior to fragment assembly? Learn how!

  6. NEBuilder_BridgingVideo_Thumb

    NEBuilder® HiFi DNA Assembly: Bridging dsDNA with a ssDNA Oligo

    Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA with a ssDNA oligo.

  7. PrimerDesignFragmentAssembly_720

    Primer Design and Fragment Assembly Using NEBuilder HiFi DNA Assembly or Gibson Assembly®

    Watch an interactive tutorial on primer design to see how simple it really is to clone with either NEBuilder HiFi DNA Assembly or the Gibson Assembly Cloning Kit.

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