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.
- Molecular Cloning Technical Guide
- 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
- Anton, B.P., Morgan, R.D., Ezraty, B., Manta, B., Barras, F., Berkmen, M. (2019) Complete genome sequence of Escherichia coli BE104, an MC4100 drivative lacking the methionine reductive pathway Microbiol Resour Announc; 8 (29), e00721-19. PubMedID: 31296691, DOI: 10.1128/MRA.00721-19
- Potapov, V., Ong, J.L., Kucera, R.B., Langhorst, B.W., Bilotti, K., Pryor, J.M., Cantor, E.J., Canton, B., Knight, T.F., Evans, T.C., Lohman, G.J.S. (2018) Comprehensive profiling of four base overhang ligation fidelity by T4 DNA ligase and application to DNA assembly ACS Synth Biol; 7 (11), PubMedID: 30335370, DOI: 10.1021/acssynbio.8b00333
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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.