DNA Ligation Kits, Ver. 1.0 and 2.1
Applications:
- High-efficiency DNA ligation in as little as 3 minutes (30 minutes for most applications)
- Suitable for λ DNA concatenation (Ver. 1)
- Ligation in small reaction volumes (Ver. 2.1)
Features:
- High efficiency DNA ligation for all cloning procedures
- Quick ligation in as little as 3 minutes (30 minutes for some applications)
- Ligation in small volumes (Ver. 2.1)
- All reagents provided
Description:
Takara's DNA Ligation Kits are simple, two-component systems that allow rapid performance of all cloning procedures requiring the covalent joining of DNA molecules in vitro. Using an optimized buffer system and T4 DNA Ligase, these kits allow ligation in 30 minutes with high transformation efficiency. For general ligation reactions, these kits allow good efficiency in only 3 minutes. The reactions can be used directly in some bacterial transformations without DNA purification.
The kits have similar, simple procedures and give nearly equivalent transformation efficiencies. However, Version 2.1 contains a convenient premix of enzyme and buffer that further minimizes pipetting steps and final reaction volume in standard reactions. Solution II of Version 2.1 is designed for linker addition ligations. Transformation efficiency can be improved by the addition of Solution III (Transformation Enhancer) into circular ligation reactions before transformation. Version 1.0 can also be used for DNA concatenation using Solution B alone.
Kit Components:
| Version 1 | |
| Solution A: Reaction Buffer | 3 x 1 mL |
| Solution B: Enzyme Solution | 2 x 187.5 μL |
| Version 2.1 | |
| Solution I: Enzyme Solution | |
| (has ligase and reaction buffer) | 3 x 250 μL |
| Solution II: Concatenation Buffer | 1 x 750 μL |
| Solution III: Transformation Enhancer | 1 x 200 μL |
| Recommended Reaction Conditions | ||
|---|---|---|
| Version 1.0 (Volume Ratio) |
Version 2.1 (Volume Ratio) |
|
| Circular DNA Ligations | ||
| Insertion of DNA fragments into plasmid vectors | DNA solution (1) | |
| Insertion of PCR fragments into T-Vectors | Solution A (4-8) | DNA solution (1) |
| Insertion of linker DNA into plasmid vectors | Solution B (1) | Solution 1 (1) |
| Self-circularization | ||
| Linear DNA Ligations | ||
| Adaptor, linker ligation to cDNA | DNA solution (1) | DNA solution (1) |
| Insertion of DNA fragments into phage vectors* | Solution B (1)(300 mM NaCl) | Solution 1 (2) Solution II (1) |
| *Version 1 is recommended | ||
References:
- Hayashi, K. et al. (1986) Nucleic Acids Res. 14:7617-31.
Figure Thumbnails:
