DNA Assembly

DNA Assembly, Cloning and Mutagenesis Kits

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.

  1. 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!

  2. 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® HiFi DNA Assembly: Bridging dsDNA with a ssDNA Oligo

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

  4. 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®.

  5. 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.

  6. How Does the NEB PCR Cloning Kit Work?

    What are toxic mini-genes, and how do they improve transformation efficiencies? Becky explains.

  7. Golden Gate Assembly Workflow

    Find out how Golden Gate Assembly can be used to quickly join multiple DNA fragments.

  8. Introduction to Gibson Assembly® E5510 E2611

    Watch an interactive tutorial that details the process by which Gibson Assembly® joins DNA fragments in a single tube, isothermal reaction.

  9. Restriction Enzymes in Golden Gate Assembly

    Type IIS restriction enzymes have both recognition and binding sites, but cut downstream of the recognition site, creating 4-base overhangs ideal for reassembly. View a list of TypeIIS enzymes.

DNA Assembly Selection Chart