RB T4 DNA Ligase
RB T4 DNA Ligase facilitates the creation of phosphodiester bonds between the 5′ phosphate terminal end and the 3′ hydroxyl group of either duplex DNA or RNA. This enzyme is versatile and can be employed for the ligation of both blunt and cohesive (sticky) ends in DNA or RNA molecules. Supplied with 10X T4 DNA ligase buffer.
Pricing
| Catalog | Description | Size | Price |
|---|---|---|---|
| RB23041 | RB T4 DNA Ligase with 10X Ligase buffer | 100 rxn | ₹ 6,500 |
| RB23042 | RB T4 DNA Ligase with 10X Ligase buffer | 500 rxn | ₹ 12,500 |
Pack Contents
RB T4 DNA Ligase
10X Ligase buffer
Nuclease free water
T4 DNA Ligase is routinely used in-house for quality control checks of all our cohesive and blunt end generating restriction enzymes. It is also used in the Blue-White standard assay.
The products are intended solely for research purposes and should not be utilized for diagnostic assays.
Store at -20°C in a constant temperature freezer
A standard ligation protocol is given below, which can be used for ligating the insert to its corresponding vector. Necessary modifications should be made as per your research needs. Ensure that the samples are prepared in sterile, DNase-free microcentrifuge tubes and incubate as recommended.
| Components | Final Concentration | 20µL reaction |
| 10X Ligation Reaction Buffer | 1X | 2 µL |
| DNA Vector * | 25ng to 100ng | Variable |
| Insert * | 1:1 to 1:10 | Variable ratio to vector |
| RB T4 DNA Ligase | 1 unit | 1 µL |
| Nuclease Free Water | – | Up to 20 µL |
*The recommended vector to insert ratio is 1:3, as it promotes the insertion of multiple fragments. However, it is recommended to try different vector to insert ratios to achieve a successful ligation. Ratios below the recommended 1:3 may reduce the ligase efficiency, but experimenting with various ratios can help optimize the ligation reaction for your specific experimental needs. Remember to prepare all samples in sterile, DNase-free microcentrifuge tubes and follow standard protocols for the ligation procedure.
The discovery and use of T4 DNA ligase in molecular biology were pivotal in advancing techniques like gene cloning and recombinant DNA technology.
The enzyme was identified in the early 1970s as part of studies on the T4 bacteriophage, which infects E. coli. This enzyme’s ability to catalyze the formation of phosphodiester bonds between DNA fragments made it essential for laboratory applications.