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.
Protocols for DNA Assembly, Cloning and Mutagenesis Kits
- KLD Enzyme Mix Reaction Protocol (M0554)
- Gibson Assembly® Electrocompetent Cells Transformation Protocol (E2611)
- Q5® Site-Directed Mutagenesis Kit (Without Competent Cells) Quick Protocol (E0552)
- Q5® Site-Directed Mutagenesis Kit (Without Competent Cells) Protocol (E0552)
- Protocol for Control Reaction (E0552)
- Transformation Protocol for Using NEB Golden Gate Assembly Mix (E1600)
- Golden Gate Assembly Protocol for Using NEB Golden Gate Assembly Mix (E1600)
- Recommended Screening Protocols for Using NEB Golden Gate Assembly Mix (E1600)
- RNA Synthesis of Cloned Insert Transcripts
- Plating Protocol for NEB PCR Cloning Kit
- Transformation Protocol for NEB PCR Cloning Kit
- Insert Screening Protocols for NEB PCR Cloning Kit
- Ligation Protocol for NEB PCR Cloning Kit
- Q5® Site-Directed Mutagenesis Kit Quick Protocol (E0554)
- Q5® Site-Directed Mutagenesis Kit Protocol (E0554)
- Protocol for Control Reaction (E0554)
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol
- NEBuilder HiFi DNA Assembly Reaction Protocol
- Ligation Protocol using BioBrick Assembly Kit® (E0546)
- Digestion Protocol for BioBrick Assembly Kit (E0546)
- NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol
- NEBuilder® HiFi Electrocompetent Transformation Protocol (E2621)
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol (E2621)
- Gibson Assembly® Chemical Transformation Protocol (E5510)
- Gibson Assembly® Protocol (E5510)
- Gibson Assembly® Master Mix – Assembly (E2611)
- Gibson Assembly® Electrocompetent Cells Transformation Protocol (E5510)
- Gibson Assembly® Chemical Transformation Protocol (E2611)
DNA Ligase Trifold
The DNA Ligase Trifold provides information about the extensive selection of DNA ligases and ligases master mixes available from NEB.
History of Cloning Poster
The History of Cloning poster presents some of the major milestones of molecular cloning technologies, in a fun format for your laboratory or classroom.
Molecular Cloning Technical Guide
Download the latest Molecular Cloning Technical Guide for help with product selection, protocols, tips for optimization and trouble-shooting.
NEBuilder HiFi DNA Assembly Bifold
Discover the next generation of gene assembly & cloning, and learn which advantages NEBuilder HiFi delivers over other gene assembly methods.
Reagents & Tools for Molecular Cloning brochure
Learn about recommended products for cloning in our Reagents and Tools for Molecular Cloning Brochure.
Synthetic biology applies the framework of engineering and circuit theory to biological questions. Learn which products and protocols you'll need to get started.
Other Tools & Resources
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 firstname.lastname@example.org.
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®.
Primer Design and Fragment Assembly Using NEBuilder® HiFi DNA Assembly or Gibson Assembly® E5510 E5520
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.
Watch an interactive tutorial that details the process by which Gibson Assembly® joins DNA fragments in a single tube, isothermal reaction.