Product Introduction

NEB recommends NEB 10-beta Competent E. coli, High Efficiency (NEB #C3019) for assemblies larger than 15 kb. NEBuilder^® HiFi DNA Assembly Bundle for Large Fragments offers a discount for purchasing these competent cells with the NEBuilder HiFi DNA Assembly Master Mix.

NEBuilder HiFi DNA Assembly Master Mix was developed to improve the efficiency and accuracy of DNA assembly. This method allows for seamless assembly of multiple DNA fragments, regardless of fragment length or end compatibility. This method has been used to assemble either single-stranded oligonucleotides or different sizes of DNA fragments with varied overlaps (15–30 bp). It has utility for the synthetic biology community, as well as those interested in one-step cloning of multiple fragments due to its ease of use, flexibility and simple master-mix format. Visit NEBuilderHifi.com for more information.

• Enjoy less screening/re-sequencing of constructs, with virtually error-free, high-fidelity assembly
• Flexible sequence design (scar-less cloning)
• No PCR clean-up step required
• Join DNA fragments together more efficiently, even with larger fragments or low DNA inputs
• Use NEBuilder HiFi in successive rounds of assembly, because it removes 5´ and 3´ end mismatches. (Save time by avoiding time-consuming PCR amplification steps.)
• Bridge two ds-fragments with a synthetic ss-DNA oligo for simple and fast construction (e.g., linker insertion or gRNA library)
• Use with lower DNA input requirements
• Easily adapted for multiple DNA manipulations, including site-directed mutagenesis
• No licensing fee requirements from NEB for NEBuilder products
• Supplied with competent cells