High-throughput cloning and automation solutions

High-throughput DNA assembly

NEBuilder^® HiFi DNA Assembly and NEBridge^® Golden Gate Assembly are leading the way in the next generation of cloning. Both methods are amenable to high-throughput workflows and scale up using automation platforms such as the Echo^® 525 Liquid Handler from Labcyte^®, Inc. and the mosquito^® LV from sptlabtech. High efficiency (> 95%) and accuracy with both cloning methods ensures you can confidently and quickly progress with accurate constructs. 

NEBuilder HiFi is recommended when working with 2–11 fragment assemblies, while NEBridge Golden Gate Assembly is appropriate for more complex designs, as well as designs containing regions of high GC content or areas of repeat. Please refer to the DNA Assembly Selection Chart for additional help determining which product is best suited for your needs.

NEBuilder HiFi DNA Assembly

• Perform less sequencing and screening of constructs with high-fidelity, virtually error-free assembly
• Enjoy compatibility with synthetic dsDNA fragments, such as gBlocks™, and ssDNA oligos
• Save time by avoiding PCR clean-up, simplifying your workflow
• Supports miniaturization with nanoliter scale volumes
  • Technical note: Modular DNA Assembly of PIK3CA Using Acoustic Liquid Transfer in Nanoliter Volumes 
  • Technical note: Miniaturized Multi-Piece DNA Assembly Using the Echo 525 Liquid Handler
  • Technical note: Nanoliter Scale DNA Assembly Utilizing the NEBuilder HiFi Cloning Kit with the Labcyte Echo 525 Liquid Handler
  • Application note: Automated high-density sample storage and miniaturized liquid handling for high-throughput synthetic biology construct assembly
• Easily adaptable to multiple site-directed mutagenesis
• Design primers and assemblies quickly and easily with our free online tool, NEBuilder Assembly Tool 

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NEBridge Golden Gate Assembly

• Experience high efficiency within regions of high GC content and areas of repeats
• Supports miniaturization
• Enjoy compatibility with synthetic dsDNA fragments, such as gBlocks
• Find flexibility with NEBridge Ligase Master Mix (NEB #M1100) and your choice of Type IIS restriction enzymes
• Design complex, high-fidelity Golden Gate Assemblies easily with our free online tools
  • NEBridge^® Golden Gate Assembly Tool – design primers, predict overhang fidelity and find optimal junctions
  • NEBridge Ligase Fidelity Tools – visualize overhang ligation preferences, predict high fidelity junction sets and split DNA sequences for scarless high-fidelity assembly

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NEB Cloning Competent Cells

• Experience high transformation efficiencies
• Enjoy compatibility with 96-well and 384-well formats
• Find the right fit with bulk formats and custom packaging options
• Choose from:
  • NEB 5-alpha Competent E. coli (High Efficiency) (NEB #C2987)
  • NEB 10-beta Competent E. coli (High Efficiency) (NEB #C3019)
  • NEB Stable Competent E. coli (High Efficiency) (NEB #C3040)
  • NEB Turbo Competent E. coli (High Efficiency) (NEB #C2984)
  • dam–/dcm^– Competent E. coli (NEB #C2925)
  • NEB 5-alpha F'I_q Competent E. coli (High Efficiency) (NEB #2992)

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High-throughput mutagenesis

Reliably create mutant libraries in a high-throughput workflow, whether making single or multiple point mutations, or performing combinatorial mutagenesis.

For single point mutations, site directed mutagenesis using Q5^® Hot Start DNA Polymerase and KLD Enzyme Mix is recommended, and for multiple sites, or combinatorial mutagenesis, we recommend NEBuilder HiFi DNA Assembly.

Single site-directed mutagenesis

• Quickly create mutant libraries with primers designed with our free online tool, NEBaseChanger^®
• Scale-up, or miniaturize with individual master mix formats compatible with automation
  • Use Q5 Hot Start High-Fidelity 2X Master Mix (NEB #M0494) for PCR amplification
  • Use KLD Enzyme Mix (NEB #M0554) for kinase, ligase and DpnI enzymatic activities in single mix
• Reduce screening of correct mutants with extremely accurate and robust Q5 High-Fidelity DNA Polymerase 
• Take advantage of automation compatible room temperature reaction set up using a Hot Start Polymerase 

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Multi site-directed mutagenesis

Using NEBuilder HiFi DNA Assembly (NEB #E2621), perform multi-site mutagenesis or combinatorial mutagenesis for diverse multi-site mutant library creation and screening. 

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High-throughput protein expression and purification

Traditional cell-based protein expression methods are challenged by limitations in speed, throughput and automation complexity. Synthesizing proteins using cell-free protein synthesis (CFPS), can overcome these shortcomings and provide a rapid and reproducible method, even for toxic proteins. CFPS components are readily dispensable by automated liquid handling devices, thus enabling high-throughput workflows and miniaturization, as described in our application note. High-throughput protein purification and separation can also be accomplished using a variety of high affinity magnetic beads and racks.

NEBExpress^® Cell-free E. coli Protein Synthesis System (NEB #E5360)

• E. coli extract-based transcription/translation engineered for high in vitro synthesis performance
• Synthesize high yields of target proteins ranging from 17 to 230 kDa (typically 0.5 mg/ml)
• Complete synthesis and visualization in approximately 2–4 hours
• Templates can be plasmid DNA, linear DNA or mRNA

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PURExpress^® In Vitro Protein Synthesis Kit (NEB #E6800)

• Reconstituted protein synthesis system includes all necessary components for in vitro transcription and translation, purified from E. coli
• Defined system with all his-tagged proteins for coupled transcription/translation; Ribosome is not his-tagged
• T7 RNA Polymerase drives in vitro transcription
• Minimal nuclease and protease activity ensure stability of synthesized protein and encoding target
• Compatible templates include plasmid DNA, linear DNA or mRNA

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NEBExpress Ni-NTA Magnetic Beads (NEB #S1423)

• An affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins in manual or automated formats
• Prepared with agarose based, super-paramagnetic microparticles which provide high binding capacity and fast magnetic response permitting high throughput and scalable purification strategies of His-tagged fusion proteins
  
  

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Need bulk sizes or a customized solution? Contact NEB's Customized Solutions Team, and we will work with you to meet your specific needs.

 

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. See www.neb.com/trademarks. The use of these products may require you to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.