The ability to ligate two or more pieces of DNA together enzymatically has found utility in many applications in the life sciences. Library preparation for Next Generation Sequencing (NGS) typically incorporates a ligation step to add bar-coded adapters to fragmented DNA, a critical step in this popular workflow. Many novel detection methodologies incorporate the ligation of DNA probes followed by a polymerase-based amplification step. For example, single nucleotide polymorphism (SNP) detection using the Ligase Chain Reaction (LCR) is a well-known application using a non-cloning thermostable ligase. Many other detection methods capitalizing on the ligation of DNA are being employed by basic and applied research groups, as well as many molecular diagnostics (MDx) companies.
Protocols for Non-Cloning Ligation
Application Notes Non-Cloning Ligation
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
Download the latest Molecular Cloning Technical Guide for help with product selection, protocols, tips for optimization and trouble-shooting.
- DNA Ligase Selection Chart
- RNA Ligase Selection Chart
- Troubleshooting Guide for Cloning
- Troubleshooting Guide for Ligases
- Troubleshooting Tips for Ligation Reactions
- Tips for Maximizing Ligation Efficiencies
Other Tools & Resources
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.