T3 DNA Ligase

T3 DNA Ligase is an ATP-dependent Ligase that catalyses blunt-end or cohesive end ligations of double-stranded DNA.

It binds to DNA at single-strand breaks and catalyzes the formation of a phosphodiester bond between adjacent 3'-hydroxyl and 5'-phosphate groups.

T3 DNA Ligase is valuable for it's ability to ligate both standard and chemically modified oligonucleotides, including those with sugar and nucleobase modifications (1). It has been demonstrated to be able to ligate xeno-nucleic acids, and has been shown to be particularly efficient with short DNA fragments (e.g. in Ligase-catalysed Oligonucleotide Polymerisation (LOOPER) reactions) in sequence directed manner (2,3).

Key Features:

• Efficient at blunt-end or cohesive end ligations
• ATP-dependent enzyme
• Salt tolerant

Applications:

• Cloning and Mutagenesis
• Ligation of standard and chemically modified oligonucleotides
• Efficient ligation of short DNA fragments (e.g. trinucleotides) e.g. LOOPER reactions

Unit Definition:

1 Unit is defined as the amount of enzyme required to ligate 50% of 1.5 µg of HindIII digested Lambda DNA at 25°C for 15 minutes in 1 x Assay Buffer

Assay Buffer:

50 mM Tris-HCl (pH 7.5 @ 25°C); 10 mM MgCl₂, 1 mM ATP, 1 mM DTT, 7.5% (v/v) Polyethylene glycol (PEG 6000)

Dilution Buffer:

10 mM Tris.HCl (pH 7.5 @ 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50 % (v/v) glycerol

We supply our T3 DNA Ligase in bulk quantities, please contact for more information.

Example Specification Sheet:

References

(1) Sabat, N., Stämpfli, A., Hanlon, S. et al. Template-dependent DNA ligation for the synthesis of modified oligonucleotides. Nat Commun 15, 8009 (2024). https://doi.org/10.1038/s41467-024-52141-8

(2) Hili R, Niu J, Liu DR. DNA ligase-mediated translation of DNA into densely functionalized nucleic acid polymers. J Am Chem Soc. 2013 Jan 9;135(1):98-101. doi: 10.1021/ja311331m. Epub 2012 Dec 26. PMID: 23256841; PMCID: PMC3544274.

(3) Khamissi N, Korfmann C, Chaudhry A, Hili R. Ligase-catalyzed transcription and reverse-transcription of XNA-containing nucleic acid polymers using T3 DNA ligase. Chem Sci. 2025 Mar 21;16(22):9749-9755. doi: 10.1039/d5sc00834d. PMID: 40160368; PMCID: PMC11951296.