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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

    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 and Mapping

    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.

    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.

    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.

    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.

    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.

    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.

    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.

    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 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.

    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.

    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.

    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.

    6 Reasons to Try RE-Mix Restriction Enzyme Master Mixes

    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?