Ligation of DNA is a central step in many modern molecular biology workflows. DNA ligases catalyze the formation of a phosphodiester bond between the 3′ hydroxyl and 5′ phosphate of adjacent DNA residues. In living organisms, ligases are essential enzymes with critical roles in DNA replication and repair. In the lab, this reaction is used to join dsDNA fragments with blunt or cohesive ends to form recombinant DNA plasmids, to add bar-coded adapters to fragmented DNA during next-generation sequencing, and to join DNA probes for SNP detection using the ligase chain reaction (LCR).
Protocols for DNA Ligation
- E. coli DNA Ligase Protocol (M0205)
- Double Digest Protocol with Standard Restriction Enzymes
- Electroporation Protocol (C2989)
- HiFi Taq DNA Ligase (M0647) Protocol
- Ligation Protocol for Cloning with Blunt/TA Ligase Master Mix (M0367)
- Ligation Protocol for Cloning with Instant Sticky-end Ligase Master Mix (M0370)
- Ligation Protocol with T3 DNA Ligase (M0317)
- Ligation Protocol with T4 DNA Ligase (M0202)
- Ligation Protocol with T7 DNA Ligase (M0318)
- NEBNext Quick Ligation Module Protocol (E6056)
- Optimizing Restriction Endonuclease Reactions
- Please see manual (NEB #E7445) for protocols
- Protocol for 9°N DNA Ligase (M0238)
- Protocol for Taq DNA Ligase (M0208)
- Protocol for ssDNA/RNA Ligation (M0319)
- Protocol for the Quick Blunting Kit (E1201)
- Removal of Single-Stranded Extension Protocol using Mung Bean Nuclease (M0250)
- Transformation Protocol (M0367)
- Transformation Protocol (M0370)
- Efficient Adaptor Ligation for the Preparation of dsDNA Libraries using the Blunt/TA Ligase Master Mix
- Joining of Difficult to Ligate dsDNA Fragments with Blunt TA Ligase Master Mix
DNA Ligase Brochure
The DNA Ligase brochure provides information about the extensive selection of DNA ligases and ligases master mixes available from NEB.
Molecular Cloning Technical Guide
The Molecular Cloning Technical Guide helps with product selection, protocols, tips for optimization and trouble-shooting.
Reagents and Tools for Molecular Cloning
Learn about recommended products for cloning in our Reagents and Tools for Molecular Cloning Brochure.
- Substrate specificity and mismatch discrimination in DNA ligases
- Properties of DNA and RNA Ligases
- Troubleshooting Guide for Cloning
- Troubleshooting Guide for Ligases
- Troubleshooting Tips for Ligation Reactions
- Tips for Maximizing Ligation Efficiencies
- Traditional Cloning Quick Guide
Other Tools & Resources
Common Applications of Exonucleases and Endonucleases
Properties of Exonucleases and Endonucleases
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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.
Ligation, the process of joining DNA fragments with a DNA ligase, proceeds in three steps. Learn more about ligation with our quick animation.