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    DNA Modifying Enzymes & Cloning Technologies

    Behind the Paper with Greg Lohman

    How can more efficient ligation of RNA-splinted ssDNA fragments improve your ligation-based detection assay? Learn about PBCV-1 DNA Ligase, also known as SplintR® Ligase.

    How do I choose the best DNA Ligase?

    NEB continues to develop and produce the most extensive commercially available selection of high-quality, and performance-optimized DNA ligases and ligase master mixes for your ligation needs.

    Why do I need to add PEG to my DNA ligation?

    Polyethylene glycol (PEG) is an important reagent in ligation reactions, find out why.

    Are some ligations more difficult than others?

    Ligation of blunt ends and single-base overhangs require optimized reaction conditions.

    What are the best conditions for DNA ligation?

    Find out how the downstream application dictates the best reaction conditions for ligation.

    What molar ratios should I use for DNA Ligation?

    The optimal reactant ratio is contingent upon the downstream application.

    Polymerases & Amplification Technologies

    Types of PCR

    NEB offers a variety of DNA polymerases for all of your experimental needs. High-fidelity, multiplex, and fast PCR are just a few of the types of amplification reactions you may need to conduct, and Breton explains which polymerases to choose.

    Tips for Amplifying Large Amplicons

    Do you need to optimize your PCR of large amplicons? Becky shares some tips for getting the most out of those long templates!

    DNA Replication with a Proofreading Polymerase

    Learn how proofreading polymerases recognize and correct mismatched bases.

    Overview of PCR Cloning

    PCR Cloning is an easy and reliable cloning method. The name is derived from the use of a DNA amplification step to generate the amplicon. Learn more about the benefits and disadvantages of PCR Cloning.

    Choose the Right DNA Polymerase for PCR

    Make sure you're using the optimal polymerase for your DNA amplifications. Get tips on choosing the right DNA Polymerase for your application.

    How to Amplify GC-rich DNA

    Looking for tips on dealing with GC-bias in DNA amplification? NEB scientists have the expertise you need!

    Why Choose Q5 High-fidelity Polymerase?

    Not sure why Q5® is your best choice for high-fidelity amplification of GC-rich targets? NEB's scientists will show you why we call Q5 an "ultra-high fidelity polymerase".

    Important Tips for Q5 Polymerase

    Here are some quick tips for getting the most out of NEB's Q5® High-Fidelity DNA Polymerase.

    Cloning & Synthetic Biology

    DNA Blunting Tutorial

    The first step in determining how your ends will be blunted is to determine if they are 5´ or 3´ overhangs. This tutorial will teach you how to identify what type of overhang you have, as well as which enzyme will blunt that end, and how.

    The Mechanism of Transformation with Competent Cells

    Transformation is the process by which bacteria are made to take up exogenous DNA. The word is derived from Griffith's discovery of a "transforming principle". Learn more about transformation and how it is used in cloning workflows.

    The Mechanism of Dephosphorylation

    Dephosphorylation is the process by which phosphate groups are removed from a molecule by a phosphatase. Removal of phosphate groups from a DNA fragment can prevent ligation. Learn more about dephosphorylation and phosphatases.

    The Mechanism of DNA Phosphorylation

    Phosphorylation is the process by which phosphate groups are added to a molecule by a kinase. The phosphorylation status of a fragment of DNA can influence its ability to proceed in reactions. Learn more about phosphorylation and kinases.

    Behind the Product: The NEB® PCR Cloning Kit

    For the inside scoop on how NEB products come to be, learn the story behind the new NEB® PCR Cloning Kit.

    How Does the NEB PCR Cloning Kit Work?

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

    Overview of PCR Cloning

    PCR Cloning is an easy and reliable cloning method. The name is derived from the use of a DNA amplification step to generate the amplicon. Learn more about the benefits and disadvantages of PCR Cloning.

    Introduction to Gibson Assembly

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

    Primer Design and Fragment Assembly Using NEBuilder HiFi DNA Assembly™ or Gibson Assembly™

    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.

    Cloning With Restriction Enzymes

    Restriction enzymes are an integral part of the cloning workflow, for generating compatible ends on fragments and vectors. This animation discusses three guidelines for determining which restriction enzymes to use in your cloning experiment.

    Overview of Traditional Cloning

    Traditional Cloning refers to the generation of DNA fragments using restriction enzymes, and their subsequent assembly and transformation. The name is derived from the method’s history as the first widely-accepted cloning method. Learn more about the benefits and disadvantages of Traditional Cloning.

    DNA Ligation

    Ligation, the process of joining DNA fragments with a DNA ligase, proceeds in three steps. Learn more about ligation with our quick animation.

    Learn How to Navigate Our New Website with Ease

    Technical Resources & Web Tools

    Search for a resource by application, product name, category, catalog number or keyword.
    Resources include:
    - Interactive tools
    - Product selection charts
    - Smartphone apps
    - Protocols and troubleshooting guides

    Powerful Search

    With NEB’s new search engine, it’s much easier to find the right product or page. Use faceted search to narrow down your results, whether it is a product or a specific FAQ, protocol or selection tool. Quickly find the exact information you are looking for.

    Streamlined Purchasing

    We designed our shopping cart with purchasing agents and real account situations in mind.
    - Email your shopping list to your purchasing agent.
    - Update account information and areas of interest to keep informed about new products or discounts.
    - Ship to multiple addresses, and keep track of them all.


    Use My NEB to customize your www.neb.com experience. Save favorite selection tools, protocols, product information or anything you visit often. Ideal for lab managers to keep a quick pick list for replenishing commonly ordered NEB products. It’s like building your own personalized catalog.

    Library Preparation for Next Generation Sequencing

    Why Choose NEBNext Reagents for NGS Library Prep?

    NEBNext products are a great choice for your NGS library preparation needs. In this video, Fiona shares a few reasons why.

    Which NEBNext Product Format Should I Choose?

    Not sure how sets and kits differ from modules? Ping explains the difference, and helps you to decide which one is right for you.

    Which DNA Library Prep Kit for Ion Torrent Should I Choose?

    Having trouble deciding which NEBNext Fast DNA Library Prep Set for Ion Torrent™ to choose? Let Christine explain which set is right for your DNA.

    Tips for Optimizing RNA Inputs in NGS Library Construction

    Watch as Deyra explains how to optimize your RNA inputs for successful NGS library preparation.

    Tips for Optimizing DNA Inputs in NGS Library Construction

    Optimizing DNA inputs for NGS library prep is an important step, if you want to ensure a high-quality library. Start by following these tips.

    Tips for Bead-based Clean ups and Size Selection

    Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

    NEBNext for Core Lab Facilities

    Are you running a core sequencing facility? Watch as Eileen shares a few reasons that NEBNext products are particularly well suited to sequencing core labs.

    12 Quick Tips for NGS Library Preparation

    Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

    Optimization of NGS Library Preparation: Low Inputs and Fast, Streamlined Workflows

    Learn how NEBNext® Ultra is enabling library prep for multiple applications, with lower input amounts and fast, streamlined workflows. In this GenomeWeb webinar Cynthia Hendrickson from HudsonAlpha Institute of Biotechnology, Momo Vuyisich from Los Alamos National Laboratory and Daniela Munafo from New England Biolabs describe use of NEBNext Ultra in exome sequencing, bacterial genome resequencing and assembly, and strand-specific RNA-Seq.

    A Breakthrough Method of RNA Sample Preparation

    The common problem of adaptor dimer formation during small RNA library construction can be avoided by using NEBNext® protocols. Learn about this technique, and how it improves both performance and sensitivity in library construction.


    Locus Specific and Global 5-hydroxymethylcytosine Detection and Quantification

    Curious about the role of 5-hydroxymethylcytosine in the genesis and function of the epigenome? Watch and learn as Sriharsa explains the study of 5-mC and 5-hmC as markers of epigenetically-modified genomic DNA, and the best methods for differentially detecting these modified species.
    How Complex is Epigenetics?

    How Complex is Epigenetics?

    Epigenetic changes to DNA impact a wide range of biological processes such as organism development and disease progression. Watch as Sriharsa enthusiastically describes this exciting and complex field that has the power to "re-write" the course of DNA transcription.
    What is Epigenetics?

    What Is Epigenetics?

    If all cells are created from the same genetic material, why are there so many different cell types? Listen to Sriharsa Pradhan, Senior Scientist, RNA Biology at NEB, as he describes how DNA is methylated and how this affects the path of reading the DNA code the same way an obstruction would derail a train off its tracks.
    How Can Epigenetics Help Us Treat Diseases?

    How Can Epigenetics Help Us Treat Diseases?

    Cancer cells have epigenetic changes to their DNA. One of these causes the turning off (silencing) of tumor suppressor genes, a small set of genes involved in cell cycle control. Sriharsa discusses drugs currently in the clinic that are able to demethylate DNA and improve the lives of cancer patients.
    Epimark Protocol Animation

    EpiMark 5-hmC and 5-mC Analysis Kit Protocol Animation

    New England Biolabs is pleased to introduce the EpiMark® 5-hmC and 5-mC Analysis Kit, a simple and robust method for the identification and quantitation of 5-hydroxymethylcytosine (5-hmC) and 5-methylcytosine (5-mC) within a specific DNA locus.

    EpiMark® is a registered trademark of New England Biolabs, Inc.

    What Does Epigenetics Have to Do with Honeybees?

    What Does Epigenetics Have to Do with Honeybees?

    Rick Feehery, Research Associate at New England Biolabs, Inc, explains the mechanisms of epigenetic DNA modification. To drive home the power of such tiny changes, he uses the example of honeybee colonies, wherein DNA methylation is the only factor that determines whether a bee is born a worker, drone or queen.
    Epigenetics Tutorial

    Interactive Tutorial Explaining the Phenomenon of Epigenetics at a Molecular Level

    Watch an interactive tutorial explaining the different molecular mechanisms by which epigenetic change influences gene expression. Learn about how NEB’s line of EpiMark® validated reagents are targeted to the various enzymes and DNA elements that are altered by epigenetic change.
    High Sensitivity 5-hmC Detection in Balb/C Brain Tissue

    High Sensitivity 5-hmC Detection in Balb/C Brain Tissue

    Follow NEB Product Development Scientist, Romas Vaisvila, Ph.D., as he demonstrates the EpiMark® 5-mc and 5-hmc Analysis Kit for locus specific identification and quantification of 5-hmc in this protocol "High Sensitivity 5-hmc detection in Balb/C Brain Tissue".

    RNA Reagents

    In Vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection

    This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene. The RNA is purified and a fraction of the uncapped RNA is enzymatically capped using the Vaccinia virus capping enzyme. In the final step, the mRNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity.

    Gene Expression & Cellular Analysis

    SNAP-tag Overview Tutorial

    View an interactive tutorial explaining the mechanism of our SNAP-tag® technologies and reagents available for researchers wishing to study the function and localization of proteins in live or fixed cells.

    Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

    Watch as Chris Provost, of New England Biolabs, performs fluorescent imaging of live COS-7 cells expressing SNAP-tag® fusion proteins.


    Overview of Glycobiology

    Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans and/or peptide portions of the glycoprotein can be accomplished with the use of deglycosylation enzymes, which are explained in detail. Unlike other chemical deglycosylation methods, enzymatic treatment is much gentler and can provide complete sugar removal with no protein degradation.

    Identification and Characterization of Protein Glycosylation

    Here we illustrate the use of glycosidases for the analysis of a model glycoprotein: recombinant human chorionic gonadotropin beta (hCGβ), which carries both N-glycans and O-glycans in this video. The technique requires only simple instrumentation and typical consumables, and it can be readily adapted to the analysis of multiple glycoprotein samples.

    Elucidating the Complexity of Heparin Oligosaccharide Analysis

    Learn more about the structural elucidation of complex and diverse heparin oligosaccharides using Bacteroides Heparinase I, II and III in combination with downstream mass spec analysis.

    Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects

    Isolating, by biopanning, the phage that binds to bone allows researchers to identify the peptide sequences that stimulate the differentiation of mesenchymal cells and potentiate bone repair.

    Restriction Enzymes

    CutSmart Restriction Enzyme Buffer

    Over 200 of NEB's restriction enzymes are 100% active in a single buffer. Learn more about CutSmart™ Buffer and why it matters to you.

    Why is My Restriction Enzyme Not Cutting DNA?

    Not getting the cleavage you expected? Let an NEB scientist help you troubleshoot your reaction.

    Restriction Enzyme Digest Problem: Too Many DNA Bands

    Are you finding unexpected bands in your digestion reaction? Here are some tips to help you determine the cause.

    Time-Saver Qualified Restriction Enzymes

    How will Time-Saver™ qualified enzymes save you time? Find out from an NEB scientist.

    Time-Saver Protocol for Restriction Enzyme Digests

    Need a protocol to digest quickly and completely? Try this protocol for Time-Saver™ qualified enzymes from NEB.

    What is Restriction Enzyme Star Activity?

    Learn what Star Activity is, why it is detrimental to accurate restriction enzyme digestion, and how NEB's HF enzymes are engineered to avoid it.

    Setting-up Restriction Enzyme Digests with RE-Mix Master Mixes

    RE-Mix® Restriction Enzyme Master Mixes offer simplified reaction setup. Learn more about digesting DNA with RE-Mix.

    RE-Mix Double Digest Protocol

    Double digests are now easier than ever! Learn how to set up your next double digest with RE-Mix®.

    Reduce Star Activity with High-Fidelity Restriction Enzymes

    NEB has engineered HF® enzymes to eliminate star activity. Learn how, and what this means for your digests.

    Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel

    Learn more about what causes this common problem, and how NEB's enzymes are QC'd to avoid DNA smearing.

    NEB Restriction Enzyme Double Digest Protocol

    Double digestions can save you time, and this video can offer tips for how to achieve the best results, no matter which of NEB's restriction enzymes you're using.

    Standard Protocol for Restriction Enzyme Digests

    Let one of NEB's restriction enzyme experts help you improve your technique and avoid common mistakes in digest setup.

    Restriction Enzyme Digest Protocol: Cutting Close to DNA End

    When cutting close to the end of a DNA molecule, make sure you know how many bases to add to the ends of your PCR primers.
    Discovering New Restriction Enzymes

    Discovering New Restriction Enzymes

    Watch as Rick Morgan, Research Scientist in the Restriction Enzyme Division, describes his passion for discovering and characterizing restriction enzymes from nature. 
    Restriction Enzyme Interaction with DNA

    The Interaction of Restriction Enzymes and DNA

    Watch as Geoff Wilson, Restriction Enzyme Division Head, describes the interaction of restriction enzymes and substrate DNA using computer models generated from x-ray crystallography data.

    6 Reasons to Try RE-Mix Restriction Enzyme Master Mixes

    Protein Expression

    What is a Disulfide Bond

    What is a disulfide bond, and how are they formed?

    Disulfide bond formation is compartmentalized (2 of 4)

    Where, and under what conditions, can disulfide bonds form?

    Disulfide bond formation in the periplasm of E. coli (3 of 4)

    What are the steps of disulfide bond formation in the periplasm, and which proteins are responsible for successful bond formation?

    Disulfide bond formation in the cytoplasm of SHuffle® cells (4 of 4)

    What makes SHuffle cells so efficient at the expression of disulfide-bonded proteins?

    Solutions for the Expression of Difficult Proteins

    NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.