DNA Assembly, Cloning and Mutagenesis Kits

Product Overview

DNA Assembly

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

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.

Site-Directed Mutagenesis

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.

Protocols for DNA Assembly, Cloning and Mutagenesis Kits

Digestion Protocol for BioBrick Assembly Kit (E0546)
Ligation Protocol using BioBrick Assembly Kit® (E0546)
Gibson Assembly® Protocol (E5510)
Gibson Assembly® Chemical Transformation Protocol (E5510)
Protocol for Control Reaction (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)
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 (NEB #M0554)
Recommended Screening Protocols for Using NEBridge Golden Gate Assembly Kit (BsaI-HFv2)
Transformation Protocol for Using NEBridge Golden Gate Assembly Kit (BsaI-HFv2) (E1601)

Application Notes for DNA Assembly, Cloning and Mutagenesis Kits

Construction of an sgRNA-Cas9 expression vector via single-stranded DNA oligo bridging of double-stranded DNA fragments
A Streamlined Cell-Free Workflow for On-Demand Protein Expression Using NEBuilder® HiFi DNA Assembly

Tools & Resources

Brochures

Golden Gate Assembly Brochure
DNA Assembly & Synthetic Biology Brochure
NEBuilder HiFi DNA Assembly Brochure
NEBuilder HiFi DNA Assembly Bifold
Molecular Cloning Technical Guide

Webinars

Quick Tips - Can I make multiple mutations with the Q5 site-directed mutagenesis kit?

Publications related to DNA Assembly, Cloning and Mutagenesis Kits

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
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
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

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. All other trademarks are the property of their respective owners. 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.

Product Listing