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T4 DNA Ligase Highlights
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the terminal 5′ phosphate and a 3′ hydroxyl group of duplex DNA, RNA, or DNA/RNA hybrids. This enzyme joins blunt and cohesive (sticky) ends, and repairs single-stranded nicks.
- Activity across a broad range of DNA and RNA inputs enables a variety of applications
- Supplied with standard or rapid ligation buffers for simple optimization
- Reduce material costs with a drop-in solution that delivers equivalent performance at a differentiated price-point
- Custom formats available, including high concentration
- Highly stringent enzyme manufacturing ensures quality performance across lots
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Applications
- Cloning of restriction digestion and PCR products
- Joining of linkers or adapters to DNA
- Nick repair
- DNA self-circularization
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High-purity
T4 Polynucleotide Kinase
Pairs seamlessly with T4 DNA Ligase for a variety of ligation applications
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QC Specifications
| Description | Specification |
|---|---|
| Protein Purity Assay | ≥ 99% |
| dsDNA Exonuclease Assay* | <1% released |
| ssDNA Exonuclease Assay* | <1% released |
| DNA contamination Assay (E. coli, mammalian, library)* | < 10 copies |
| Phosphatase Contamination Assay* | < 1% released |
| Endonuclease Contamination Assay* | Not detectable |
*As assessed using 6,000 U of enzyme input per assay.
Properties
Unit definition: One unit is defined as the amount of DNA ligase required to join 50% of 100 ng of DNA fragments with cohesive termini in 20 μL 1X DNA Ligase Buffer following a 30-minute incubation at 23°C
Reaction conditions:
1X Rapid Ligation Buffer*
Incubate at 20-25°C
Storage buffer: 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol pH 7.5 @ 25°C
1X Rapid Ligation Buffer: 6% PEG, 66 mM Tris-HCl, 10 mM MgCl₂, 1 mM DTT, 1 mM ATP pH 7.6 @ 25°C
Heat inactivation: 65°C for 10 min
Molecular weight: 55.3 kDa
*Note: Other buffer formulations are also available