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LET NEB’S SCIENTIFICEXPERTISE HELP DRIVE YOURINNOVATIONS FORWARDIn recent years, there has been a dramatic risein the number of biologics and companiondiagnostics developed and commercialized, andthese important areas of biotechnology are morereliant than ever on cutting edge molecular biologytools and techniques. For instance, companiondiagnostics, based on DNA amplification, canfoster a better match of effective therapies withpatients; personalized medicines, utilizing nextgeneration sequencing, can also help tailortreatments for specific patients; and a wide rangeof biologics is now being developed as a result ofadvancements in recombinant technologies.2

COLLABORATIONWe’re here to champion your efforts. Wehave extensive scientific expertise and candevote the necessary attention to developingsolutions for your specific needs – alongwith the manufacturing capacity to scale upquickly. Whether you’re looking for a customversion of an existing product, to find a newway around a development roadblock, or tolicense one of our technologies, our team isready to work with you.PRODUCT PORTFOLIOTHE NEB DIFFERENCEAt NEB, we have decades of experiencein practicing molecular biology, which hasled to the introduction of a broad productportfolio that has the potential to touchalmost every stage of today’s biotechnologydiscovery and development processes. Withthis expertise in hand, we are ready and ableto collaborate with biotech companies,large and small, to support their discoveriesand help them move through clinical testing,manufacturing, and quality control, and intoregulated markets around the world.We can provide an entire suite of enzymesthat have the potential to accelerateyour discovery efforts. Our expertise inenzymology has enabled us to develop uniqueenzymes that enable faster, more robustworkflows. Further, enzymes can be providedboth in small aliquots and in bulk, in differentformats (liquid, lyophilized and glycerol-free),as well as packaged into complete kits. And ifyou cannot find what you need, let’s see if wecan come up with a solution that does!QUALITYOur attention to quality is second to none.We are fully compliant and certified to ISO9001:2015 and ISO 13485:2016 standards,and have recently completed building a GMPmanufacturing facility for reagents used inmore regulated markets.TECHNICAL SUPPORTYou can be sure that the people you contactfor technical support are as knowledgeable asthe scientists who work in our labs – becausethey’re one and the same. When you pick upthe phone or send an email, you get rightto the source of useful information andeffective solutions.GLOBAL SUPPLY CHAINIn addition to our headquarters in the USA,NEB has warehousing capabilities in Canada,China, UK, Germany, Japan and Singapore,which can be used to not only access ourproducts rapidly, but also get them to yourcollaborators and customers. NEB also hasa number of Freezer Program options thatcombine onsite access with improved ways ofmanaging your lab inventory and cash flow.3

THINK OF NEB AS YOUR EXPERIENCEDPARTNER THROUGHOUT THEBIOTECHNOLOGY DISCOVERY PROCESSSYNTHETIC BIOLOGY AND DNA ASSEMBLYA more recent expansion of the biotechnology field, synthetic biology utilizesgenes and proteins as parts or devices, with the goal of re-designing and/orassembling these parts in novel ways to create a new and useful functionality.Recent advances in biofuels generation, the production of biochemicals,and understanding the minimal genome all benefit from synthetic biologicalapproaches. Historically, conventional genome engineering in the agriculturalindustry has involved the insertion of new genes in plants. Synthetic biologyapproaches enable the insertion of entire genetic pathways to producedesirable traits, such as disease resistance or improved nutritional value.NEB provides a widerange of products tosupport synthetic biology, DNAassembly and other cloningtechniques. Find out how theseproducts can speed up yourworkflows for both simple andcomplex reaction design atCloneWithNEB.com.The pharmaceutical industry has also benefitted from advances in syntheticbiology. Examples include the design of minimal synthetic cells, containinga genetic circuit linking genetic regulators with an output signal, such asluminescence or fluorescence for phenotypic cell-based screening, thegeneration of small molecule libraries from the reorganization of syntheticbiology parts, and cell-free metabolic engineering.Often these projects rely on the ordered assembly of multiple DNA sequencesto create large, artificial DNA structures. To this end, methods have evolved tosimplify this process. Tools such as NEBuilder HiFi DNA Assembly(NEB #E2621, E5520, E2623) and NEB Golden Gate Assembly Kits(NEB #E1601, E1602) can be used to rapidly create many functional DNAstructures, from a simple joining of two metabolic genes, all the way up to thecreation of an artificial genome.FEATURED PRODUCT:NEBuilder HiFi DNA Assembly Not your average DNA assembly reagent2-fragment assemblyssOligo & dsDNA assembly4-fragment assemblyAnnealed-oligo assemblyNEB #E2621S/L/XE njoy simple and fast seamless cloning in as littleas 15 minutesU se one system for both "standard-size" cloningand larger gene assembly products, up to12 fragmentsD NA can be used immediately for transformation oras template for PCR or RCAA dapts easily for multiple DNA manipulations,including site-directed mutagenesis15-bp overlap25-bp overlapSticky endHigh efficiency3 - and 5 -end mismatch assemblySite-directed mutagenesisMultiple sitesD esign primers using our free online tool atNEBuilder.neb.comN o licensing fee requirements from NEB forNEBuilder products4Visit NEBuilderHiFi.com for more details.Blunt endNEBuilder HiFi DNA Assembly can be used for a variety of DNA assembly methods.

Perhaps no other tool in genomic engineering has receivedmore attention of late than CRISPR-based methods.Programmable nucleases enable the generation ofpermanent mutations by generating site-specific doublestranded breaks. Knock-outs and knock-ins can be rapidlyintroduced in cell lines and model organisms.The validation of small molecule targets by knocking outgene function or rescuing drug sensitivity by modifyingdrug resistant alleles is now a standard method for targetvalidation. By combining next generation sequencing for thediscovery of compound-resistant mutations and CRISPRbased methods for target validation, drug-target pairs canbe identified and validated in model systems, such asmammalian cells, as well as model organisms.NEB provides reagents to support a broad variety ofCRISPR/Cas9 genome editing approaches. From theintroduction of Cas9 and single guide RNA (sgRNA) intoplasmids, to direct introduction of Cas9 ribonucleoprotein(RNP), and the detection of edits using next generationsequencing or enzymatic mutation detection, NEBprovides reagents that simplify and shorten genome editingworkflows.For more details on how NEB can helpimprove your genome editing efficiencies,visit www.neb.com/GenomeEditing.FEATURED PRODUCT:EnGen Cas9 NLS NEB #M0646T/M Ideal for direct introduction of Cas9/sgRNA complexes Dual NLS for improved transport to the nucleus ompatible with the EnGen sgRNA Synthesis Kit,CS. pyogenes (NEB #E3322S) and the EnGen MutationDetection Kit (NEB #E3321S)Increased genome editing efficiency usingCas9 RNP delivery10080% modificationGENOME EDITING (CRISPR/CAS9)6040200Untransfected PlasmidDNAmRNACas9EnGenCas9 NLSCas9 and sgRNA targeting a human gene were delivered to HEK293 cells bytransfection. Transfected plasmid DNA contained expression cassettes for 2x NLS(N- and C-terminal) Cas9 and sgRNA. Plasmid DNA was delivered using TransIT-X2(Mirus). Transfected mRNA was modified with pseudouridine and 5-methylcytosineand encoded 2x NLS (N-and C-terminal) Cas9. sgRNA was co-transfected with themRNA using TransIT-mRNA. Cas9 RNPs were delivered in reverse transfections usingLipofectamine RNAiMAX (Life Technologies) using 10 nanomolar final concentrationof ribonucleoprotein (RNP). Cas9 has no NLS in the protein sequence. EnGen Cas9has N- and C-terminal NLSs. The efficiency of editing was determined using T7Endonuclease I assay and is expressed as % modification.Visit www.neb.com/M0646 for more details.5

Next generation sequencing (NGS) has proven a valuabletechnology across a broad range of activities involvedin the development and application of novel therapies.Library preparation is a critical part of the NGS workflow;successful sequencing requires the generation of highquality libraries of sufficient yield and quality. Further,with sample numbers and types increasing, highperformance is required from ever-decreasing inputquantities and from samples of lower quality.With over 40 years of expertise in enzyme researchand manufacturing, NEB is a world leader in thedevelopment of innovative technologies supportingsample prep for next generation sequencing. Withover 70 application-specific products available, theNEBNext product portfolio supports a broad rangeof sequencing based technologies, including wholegenome sequencing, transcriptome and single-cellsequencing (RNA-seq), and target enrichment.FEATURED PRODUCT:NEBNext Ultra II FS DNALIbrary Prep Kit for Illumina NEB #E7805S/L erform fragmentation, end repair and dA-tailing in a singlePenzyme mix xperience reliable fragmentation with a single protocol,Eregardless of DNA input amount or GC content repare high yields of high quality libraries from a wide rangePof input amounts: 100 pg – 0.5 µgNEBNext Ultra II FS DNA produces the highest yields,from a range of input amounts140120NEBNext Ultra II FS Kapa HyperPlusCovaris Illumina Nextera Library Yield (nM)NEXT GENERATION SEQUENCING100 806040200PCR cyclesDNA inputFind out how NEBNext kits can helpstreamline your workflows and generatehigh quality libraries, even with limited amountsof DNA or RNA, at NEBNext.com.613100 pg10500 pg91 ng550 ng4100 ng3500 ngLibraries were prepared from Human NA19240 genomic DNA. For NEBNext Ultra II FS, a20-min. fragmentation time was used. For Kapa HyperPlus libraries, input DNA was cleaned upwith 3X beads prior to library construction, as recommended, and a 20-min. fragmentation timewas used. Illumina recommends 50 ng input for Nextera, and not an input range; therefore,only 50 ng was used in this experiment. “Covaris” libraries were prepared by shearing in 1X TEBuffer to an insert size of 200 bp using a Covaris instrument, followed by library constructionusing the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Error bars indicate standarddeviation for an average of 3–6 replicates performed by 2 independent users.Visit www.neb.com/E7805 for more details.

NOVEL SOLUTIONS TO ADVANCEYOUR TECHNOLOGIES FASTERTARGET ENRICHMENTGenomic information has been widely applied to the development of novel biomarkersassociated with a wide variety of human diseases. Techniques including whole genomesequencing, whole exome sequencing and RNA sequencing are now routinely applied toidentifying genetic variation across patient cohorts demonstrating common phenotypesin the presentation of disease. These data are used for the identification of therapeutictargets, as well as the elucidation of disease pathways and mechanisms of therapeuticresistance. Downstream in these pipelines, genomic data is increasingly being applied topharmacogenomics studies aimed at understanding the efficacy of therapies on stratifiedpatient populations.Inclusion of genomic profiling in clinical trials has become commonplace, bothprospectively and retrospectively. For prospective studies, targeted gene panels arebeing applied across trial participants, with the goal of matching therapeutic responseoutcomes with patient genotypes. Retrospectively, broad genomic profiling of patientswith either a positive or negative therapeutic response are being used to identify specificvariants with the potential to serve as biomarkers to predict therapeutic efficacy.Finally, targeted gene panels focusing on specific biomarkers with proven predictivevalue are being developed as companion diagnostics. These assays, which are includedas part of a therapy’s FDA approval process, are now commonly used to determinetherapeutic applicability for a specific patient.Regardless ofsample type orassay content, NEBNextDirect allows you toenrich your targets withprecision. Learn more atNEBNextDirect.com.NEBNext Direct employs a unique technology that enables highly specific targetenrichment of genomic regions of interest. This innovative approach balances thespeed and precision of multiplexed PCR-based approaches with the content scalabilitytypical of hybridization-based methods. This flexibility allows a single workflow for assaysranging from single gene tests to comprehensive panels including several hundred genes.FEATURED PRODUCT:NEBNext DirectCustom Ready Panels NEBNext Direct Custom Ready Panelsdemonstrate optimum performance across awide range of panel sizes1 gene (3 kb)10 gene (30 kb)NEB #E6631S/L/XE xperience unmatched specificity andcoverage uniformityE liminate synthesis and optimization steps forfaster turnaroundI mprove sensitivity with our Unique MolecularIndex (UMI)G enerate results in one day with ourautomation-friendly workflowVisit NEBNextDirect.com for more details.Percent (%)C hoose from a single gene to hundreds of genes1009080706050403020100% Aligned25 gene (70 kb)50 gene (150 kb)% Selected100 gene (280 kb) 25% ofMean CoverageKey target enrichment metrics demonstrate consistent performance across a range ofpanel sizes. 100 ng of DNA was tested against panels of 1, 10, 25, 50 and 100 genes,and sequenced using Illumina paired-end 150 bp sequencing. Larger panels included allgenes present in smaller panels.For research use only; not intended for diagnostic use.7

UNIQUE SOLUTIONS TOSTREAMLINE YOUR WORKFLOWSAND MAXIMIZE YIELDSIN VITRO TRANSCRIPTIONNEB offers aselection of HiScribeRNA synthesis kits for thegeneration of high yields ofhigh quality RNA that canbe used in a wide variety ofapplications. Find out howNEB can streamline your RNAworkflows at NEBrna.com.mRNA-based medicine holds great promise for applications in infectious diseasevaccination, cancer vaccination, as well as other disease treatments. Deliveryof mRNA is thought to present less risk than DNA-based therapies, and hasadditional benefits in speed of production and personalization, versus proteindelivery. Functional mRNA requires the presence of a m7G cap and poly(A) tail forrecognition by the host cellular translation system. In addition, other structuresmay be desirable, such as the presence of modified bases or Cap-1 structures.NEB provides a variety of tools for the in vitro transcription (IVT) of RNAand mRNA. Reagents can be purchased separately or as optimized kits. Thesereagents support applications ranging from benchtop research to large scale RNAproduction and purification for clinical applications.NEB also provides kits for fast and simple cleanup or synthesized RNA.Monarch RNA Cleanup Kits ( NEB #T2030, T2040, T2050) are available forpurification of 10, 50 and 500 µg of RNA, and efficiently removes reactioncomponents, including unincorporated nucleotides, resulting in highly pure RNAfor downstream applications.Looking for GMPgrade* reagents formanufacturing therapeuticRNA? Visit www.neb.com/GMP to find out how NEBcan help.* “GMP-grade” is a branding term NEB uses to describe reagents manufactured at NEB’s Rowley facility. The Rowley facility was designedto manufacture reagents under more rigorous infrastructure and process controls to achieve more stringent product specifications andcustomer requirements. Reagents manufactured at NEB’s Rowley facility are manufactured in compliance with ISO 9001 and ISO 13485quality management system standards. However, at this time, NEB does not manufacture or sell products known as Active PharmaceuticalIngredients (APIs), nor does NEB manufacture its products in compliance with all of the Current Good Manufacturing Practice regulations.FEATURED PRODUCT:HiScribe T7 High Yield RNASynthesis KitRobust RNA Synthesis from a variety of template sizesusing the HiScribe T7 High Yield RNA Synthesis Kit Up to 180 μg of RNA per reaction from 1 μg of control templateE nables full substitution of NTPs for labeling and incorporationof modified basesL inearized control template included for verification of RNAsynthesisµg RNA/ReactionNEB #E2040S2001801601401201008060402000.3 kb0.8 kb1.8 kb020406080100Reaction time, minutes120140Time course of standard RNA synthesis from three DNA templates of different sizes usingHiScribe T7 High Yield RNA Synthesis Kit. Reactions were assembled according to theprotocol and incubated at 37 C for the indicated time. Transcripts were purified over spincolumns and quantified on a NanoDrop Spectrophotometer to calculate reaction yield.Visit www.neb.com/E2040 for more details.8

PROTEIN EXPRESSION &PURIFICATIONRecombinant production of proteins is one of themost powerful techniques used in biotechnology.The ability to produce an abundance of a desiredprotein can enable a wide range of possibilities,including its use in industrial processes or todiagnose or treat disease.At first glance, recombinant protein expressionlooks quite simple. However, it can be verychallenging, because so many factors influencethe process. For example, each protein foldsin its own unique manner; some proteins requirepost-translational modifications, and someproteins have activities that are detrimental totheir host. Thus, no single solution exists forthe successful production of all recombinantproteins.FEATURED PRODUCT:PURExpress In VitroProtein Synthesis Kit NEB #E6800S/L Express a wide range of proteins free of modifications and degradation Save valuable time with results available in just a few hoursUse with plasmid DNA, linear DNA or mRNA One-step reaction requiring the mixing of only two tubesProtein expression using the PURExpress In Vitro ProteinSynthesis -100 —80 —60 —50 —Focused on prokaryotic systems, NEB wasone of the first companies to recognize thepromise of recombinant technologies, andhas since developed a wide array of proteinexpression solutions, including many forproteins that are difficult to express. Each NEBtechnology offers different advantages, whichenable you to choose the strategy that best suitsyour protein expression and purification needs.Visit www.neb.com/ProteinExpressionto view NEB’s full portfolio of products.40 —30 —25 —20 —15 —10 —Reactions were carried out according to manual recommendations. Red dot indicates protein of interest.Marker M is the Protein Ladder (NEB #P7703).Also available: NEBExpress Cell-free E. coli ProteinSynthesis Kit. For more details on these products, visitwww.neb.com/ProteinExpression.9

GLYCOPROTEOMICSOver the past 20 years, the list of enzymes availablefor glycan analysis has steadily grown, and manyof these enzymes have transitioned from academicresearch into workflows that both characterizeglycans on biotherapeutics and support glycanbased clinical tests. As a result of this, and thegrowing importance and understanding of proteinglycosylation in disease, new demands are beingplaced on the quality of reagents used for glycananalysis.NEB is committed to continually improvingthe quality of its glycosidases with the aim ofsupporting their routine use in biopharma andclinical diagnostic applications. Many of thesereagents are recombinant, and all undergo severalquality control assays, enabling us to provideproducts with lower unit cost, higher purity, andreduced lot-to-lot variation.Visit NEBglycosidase.com to learnmore, view the full list of productsavailable, and to download our white paper,“Redesigning glycosidase manufacturing qualityfor pharmaceutical and clinical applications.”FEATURED PRODUCT:Rapid PNGase FNEB #P0710S Speed up deglycosylation with one-step reaction setupRelease all N-glycans without bias chieve sensitive and reproducible results with shortAreaction times Move directly to downstream analysisRapid PNGase F results in complete deglycosylationwith a short incubationRapid PNGase F (5 min. at 50 C)DeglycosylatedIgGA monoclonal murine antibody was incubated for 5 min. with Rapid PNGase F,demonstrating that a short incubation is sufficient to deglycosylate every N-glycansite without bias.Visit www.neb.com/PNGaseF for more details.10

APPLY NEB’S SCIENTIFIC ANDMANUFACTURING EXPERIENCETO HELP DRIVE YOUR INNOVATIONARE YOU READY TO PARTNER WITH NEB?LET’S GET THE CONVERSATION STARTED! Visit www.neb.com/DriveYourInnovation to learn more and request a discussion or visit from your NEB representative Learn more about custom or bulk opportunities at www.neb.com/CustomizedSolutions Starting a new laboratory or company? Contact your local NEB representative to see how we can help you!For international customers, visit www.neb.com/internationalorders to locate your local NEB point of contact.Developing a newlife science productor technology?CONTACT OUR CUSTOMIZEDSOLUTIONS TEAM TO STARTBUILDING YOUR SOLUTION.There are multiple considerations when selecting a reagentpartner for your custom needs – technical expertise, scale,turnaround time, quality, and logistical support are all criticalto meeting your goals. And if you are working in a regulatedmarket, that list of concerns and hurdles increases considerably. At NEB, we have taken these factors into accountand can provide you with a unique solution that allows you toaccess novel products, meet quality specifications, speed time to market, andultimately streamline your supply chain – leaving you more time for innovation.NEB has over 45 years of experience in the discovery, development and manufacture of molecularbiology reagents. These are essential components in a vast array of genomic and proteomic technologies that continue to transform our understanding of the world we live in, and ultimately the diagnosisand treatment of disease. Our Customized Solutions Team was purpose-built to leverage what we dowell. Our uniquely collaborative spirit, commitment to quality, scale of manufacturing and exceptionaltechnical and customer support all help to ensure the success of our partners.Contact us at custom@neb.com or visit www.neb.com/CustomizedSolutions to learn more.11

GERMANY & AUSTRIANew England Biolabs GmbHBrüningstr. 50, Geb B85265926 Frankfurt/Main, GermanyFax: 49/(0)69/305-23149Free Call: 0800/246 5227 (Germany)Free Call: 00800/246 52277 (Austria)info.de@neb.comFRANCENew England Biolabs France SASGenopole Campus 1, Bâtiment 65 rue Henri Desbruères91030 Evry cedex, FranceTel.: 0800 100 632 (Customer Service)Tel.: 0800 100 633 (Technical line.frHEADQUARTERS:USANew England Biolabs, Inc.info@neb.comwww.neb.comFor a complete list of all NEB subsidiariesand distributors, please go r local NEB comITALY:EUROCLONE omBULGARIAELTA 90M Ltd.office@elta90.euwww.elta90.comISRAEL:Ornat BioChemicals &Lab Equipmentornatbio@ornat.co.ilwww.ornat.co.ilSLOVAK REPUBLICBioTech ostica Skalpeli d.o.o.skalpeli@skalpeli.hrwww.skalpeli.hrCZECH REPUBLIC:BioTech ika Denmark ordika E litex Kft.info@kvalitex.huwww.kvalitex.huISO 14001ISO al DevicesRegisteredPOLAND:Lab-JOT Ltd. Sp.z o.o. ontact@biozyme.rowww.biozyme.roRUSSIA:SkyGen LLCinfo@skygen.comwww.skygen.comSERBIA:Alfa Genetics d.o.o.office@alfagenetics.rswww.alfagenetics.rsISO 9001RegisteredNORWAY:BioNordika TECH – Version 2.0 – 05/20x/f/yNEB129SLOVENIA:Mikro Polo d.o.o.info@mikro-polo.siwww.mikro-polo.comSOUTH AFRICA:Inqaba Biotechnical Industries(Pty) EDEN, ESTONIA, LATVIA,LUTHUANIA:BIONORDIKA SWEDEN ioConcept Ltd.info@bioconcept.chwww.bioconcept.chTURKEY:EKA BIOLAB TEKNOLOJIEmail: info@ekabiolab.comwww.ekabiolab.comVisit neb.com to viewour Corporate SocialResponsibility &Sustainability BrochureIn this brochure, you’ll learn more about NEB’scommitment to environmental stewardship,scientific advancement and education, humanitarianefforts and employee well-being.

based methods for target validation, drug-target pairs can be identified and validated in model systems, such as mammalian cells, as well as model organisms. NEB provides reagents to support a broad variety of CRISPR/Cas9 genome editing approaches. From the introduction of Cas9 and single guide RNA (sgRNA) into

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