| CONFIGURATION | |
|---|---|
| Kit contents | T4 DNA Ligase (120 U/µL or 600 U/µL). Configurations also include: 10X Ligation Buffer, 5X Rapid Ligation Buffer or 2X Rapid Ligation Buffer |
| 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 |
| Enzyme storage buffer | 20 mM HEPES-KOH, 0.1 mM EDTA, 50 mM KCl, 8 mM DTT, 50% glycerol, pH 7.6 |
| 10X Ligation Buffer: | 500 mM Tris-HCl, 100 mM MgCl₂, 50 mM DTT, 1 mM ATP pH 7.6 @ 25°C |
| 5X Ligation Buffer: | 30% PEG, 330 mM Tris-HCl, 50 mM MgCl₂, 5 mM DTT, 5 mM ATP pH 7.6 @ 25°C |
| 2X Ligation Buffer: | 15% PEG, 132 mM Tris-HCl, 20 mM MgCl₂, 2 mM DTT, 2 mM ATP pH 7.6 @ 25°C |
| QUALITY CONTROL | |
|---|---|
| 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.
| SHIPPING AND HANDLING | |
|---|---|
| Shipping conditions | Ice packs |
| Storage | -20°C ± 5°C |
| Shelf life (off-the-shelf products) | ≥ 12 months |
| Shelf life (custom products) | ≥ 18 months |