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| Product Name | T4 DNA Ligase (5 U/μL) |
| Catalog No. | IVDRM-0101 |
| Description | T4 DNA Ligase (5 U/μL) is a high-quality ligase that, using ATP as a cofactor, catalyzes the formation of phosphodiester bonds between adjacent 5'-phosphate and 3'-hydroxyl termini in double-stranded DNA with either blunt or cohesive ends. It can also catalyze the joining of RNA to single-stranded DNA or RNA strands within a duplex, but it does not ligate fully single-stranded nucleic acids. With no detectable exonuclease or nicking activity and excellent ligation efficiency, this enzyme is well suited for RNA 3'-end labeling, circularization of RNA and DNA oligonucleotides, cDNA cloning, and other nucleic acid manipulations. |
| Intended Use | RNA 3'-end labeling, circularization of RNA and DNA oligonucleotides, cDNA cloning, and general nucleic acid ligation. |
| Principle / Technology | In the presence of ATP as a cofactor, T4 DNA Ligase catalyzes the formation of phosphodiester bonds between the 5'-phosphate and 3'-hydroxyl ends of adjacent nucleotides in double-stranded DNA (blunt or cohesive ends). It also joins RNA to ssDNA or RNA strands hybridized in a duplex, but cannot ligate fully single-stranded nucleic acids. |
| Sample Type | Double-stranded DNA (blunt or cohesive ends), RNA/DNA oligonucleotides, RNA in duplex structures. |
| Performance Range / Specifications | Concentration: 5 U/μL High ligation efficiency No detectable exonuclease or nicking activity |
| Specificity | Ligates adjacent 5'-phosphate/3'-hydroxyl termini in duplex structures; does not catalyze ligation between fully single-stranded nucleic acids. |
| Reaction Conditions / Protocol | Perform ligation with ATP-containing ligation buffer according to standard protocols for blunt-end or cohesive-end ligation. |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 2 years when stored at -20°C. |
| Package Specifications | 1,000 U 50,000 U |
| Quality Control | Tested to be free of exonuclease and nicking activity; ligation efficiency verified. |
| Purity | Free of detectable exonuclease and nicking enzyme contamination. |
| Concentration | 5 U/μL |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 2 years at -20°C. |
| Compatibility | Suitable for vector-insert ligation (including dephosphorylated vectors; the resulting nick at the insert 3' end is repaired in E. coli) and compatible with standard NGS adapter ligation workflows. Note: reactions containing ligation enhancer with high PEG content must be purified before fragment size selection. |
| Recommended Buffer System | ATP-containing T4 DNA ligase reaction buffer. |
| Application Notes / Precautions | When ligating inserts into dephosphorylated vectors, a nick remains between the insert 3' end and the vector 5' end and is repaired automatically in E. coli. If a ligation enhancer containing high concentrations of PEG is used, the ligation product must be purified before fragment size selection, as PEG significantly interferes with sorting. |
For research use only, not for clinical use.
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