The successful assembly of DNA can be utilized for both cloning and synthetic biology (in which genes and proteins are redesigned and/or assembled into novel ways to create new and useful functionality). New England Biolabs offers several products that can be used for DNA assembly and cloning of multiple DNA fragments, including NEB Gibson Assembly®, NEBuilder® HiFi DNA Assembly, which utilizes a higher fidelity polymerase, and Golden Gate Assembly. Please refer to the DNA Assembly Selection Chart to determine which product would work best for your needs.
PCR cloning is a rapid method for cloning genes, and is often used for projects that require higher throughput than traditional cloning methods can accommodate. It also allows for cloning of fragments that are not available in large amounts. NEB offers an affordable cloning kit as an alternative to TOPO®. The NEB PCR Cloning Kit allows quick and simple cloning of all your PCR amplicons, regardless of the polymerase used. This kit utilizes a novel mechanism for background colony suppression – a toxic minigene is generated when the vector closes upon itself – and allows for direct cloning from your PCR reaction with no purification step. The NEB PCR Cloning Kit is available with and without competent cells.
NEB offers the Q5 Site-Directed Mutagenesis Kit as an alternative to QuikChange™. The kit allows for rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hours. The Q5 Site-Directed Mutagenesis Kit is available with and without competent cells.
- Digestion Protocol for BioBrick Assembly Kit (E0546)
- Ligation Protocol using BioBrick Assembly Kit® (E0546)
- Gibson Assembly® Master Mix – Assembly (E2611)
- Gibson Assembly® Protocol (E5510)
- Gibson Assembly® Chemical Transformation Protocol (E5510)
- Protocol for Control Reaction (E0554)
- Q5® Site-Directed Mutagenesis Kit Protocol (E0554)
- Q5® Site-Directed Mutagenesis Kit Quick Protocol (E0554)
- Gibson Assembly® Chemical Transformation Protocol (E2611)
- Gibson Assembly® Electrocompetent Transformation Protocol (E2611)
- Gibson Assembly® Electrocompetent Transformation Protocol (E5510)
- Insert Screening Protocols for NEB PCR Cloning Kit
- Ligation Protocol for NEB PCR Cloning Kit
- Plating Protocol for NEB PCR Cloning Kit
- Transformation Protocol for NEB PCR Cloning Kit
- Protocol for Control Reaction (E0552)
- Q5® Site-Directed Mutagenesis Kit (Without Competent Cells) Protocol (E0552)
- Q5® Site-Directed Mutagenesis Kit (Without Competent Cells) Quick Protocol (E0552)
- NEBuilder HiFi DNA Assembly Reaction Protocol
- NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol (E2621, E5520, E2623)
- Golden Gate Assembly Protocol for Using NEB Golden Gate Assembly Mix (E1600)
- Recommended Screening Protocols for Using NEB Golden Gate Assembly Mix (E1600)
- Transformation Protocol for Using NEB Golden Gate Assembly Mix (E1600)
- RNA Synthesis of Cloned Insert Transcripts
- KLD Enzyme Mix Reaction Protocol (M0554)
- Golden Gate Assembly Protocol for Using NEBridge Golden Gate Assembly Kit (BsaI-HFv2) (E1601)
- Recommended Screening Protocols for Using NEBridge Golden Gate Assembly Kit (BsaI-HFv2)
- Transformation Protocol for Using NEBridge Golden Gate Assembly Kit (BsaI-HFv2) (E1601)
- DNA Assembly & Synthetic Biology
- Golden Gate Assembly brochure
- History of Cloning Poster
- Molecular Cloning Technical Guide
- NEBuilder HiFi DNA Assembly Bifold
- Quick Tips - Can I make multiple mutations with the Q5 site-directed mutagenesis kit?
- Prediction of Golden Gate Assembly GGA Using a Comprehensive Analysis of T4 DNA Ligase End-Joining Fidelity and Bias (2018)
- 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
- Carpinone, E.M., Li, Z., Mills, M.K., Foltz, C., Brannon, E.R., Carlow, C.K.S., Starai, V.J. (2018) Identification of putative effectors of the Type IV secretion system from the Wolbachia endosymbiont of Brugia malayi PLoS One; 13 (9), e0204736. PubMedID: 30261054, DOI: 10.1371/journal.pone.0204736
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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.
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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.
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!
Learn how you can use NEBuilder HiFi to generate an sgRNA-Cas9 expression vector with a single-stranded oligo bridge.
Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA with a ssDNA oligo.
Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB 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.
What are toxic mini-genes, and how do they improve transformation efficiencies? Becky explains.
Find out how Golden Gate Assembly can be used to quickly join multiple DNA fragments.
This video demonstrates how to use the Golden Gate Assembly Tool, we will walk through selecting insert and plasmids, primer design to make amplicon inserts.
Watch an interactive tutorial that details the process by which Gibson Assembly® joins DNA fragments in a single tube, isothermal reaction.