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DNA Ligation

Ligation of DNA is a critical step in many modern molecular biology workflows. The sealing of nicks between adjacent residues of a single-strand break on a double-strand substrate and the joining of double-strand breaks are enzymatically catalyzed by DNA ligases. The formation of a phosphodiester bond between the 3' hydroxyl and 5' phosphate of adjacent DNA residues proceeds in three steps: Initially, the ligase is self-adenylated by reaction with free ATP. Next, the adenyl group is transferred to the 5'-phosphorylated end of the "donor" strand. Lastly, the formation of the phosphodiester bond proceeds after reaction of the adenylated donor end with the adjacent 3' hydroxyl acceptor and the release of AMP. In living organisms, DNA ligases are essential enzymes with critical roles in DNA replication and repair. In the lab, DNA ligation is performed for both cloning and non-cloning applications.

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DNA Ligation includes these areas of focus:
Non-Cloning Ligation
Cloning Ligation
Protocols for DNA Ligation
Application Notes DNA Ligation
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This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Videos

  1. DNA Ligase

    DNA Ligase Fidelity: when does it matter?

    High fidelity polymerases are everywhere—but why would you need a high fidelity ligase? And what do we even mean by “fidelity” when we’re talking about ligation? In this webinar, NEB Scientist and ligase expert Greg Lohman discusses mismatch ligation by DNA ligases and the molecular diagnostics applications that depend on the use of high-fidelity DNA ligases like NEB’s HiFi Taq DNA Ligase to detect single base differences in DNA.

  2. DNA Ligase

    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.

  3. WhichLigase_thumb

    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.

  4. BestRatios_thumb

    What molar ratios should I use for DNA Ligation?

    The optimal reactant ratio is contingent upon the downstream application.

  5. WhyUsePEG_thumb

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

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

  6. BestReactionTimes_thumb

    What are the best conditions for DNA ligation?

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

  7. DifficultLigation_thumb

    Are some ligations more difficult than others?

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

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