This document introduces the product information of DNA Ligation Kit, including the product's usage methods, storage conditions, and precautions.
This document provides instructions for using the Agilent DNA Ligation Kit, which contains reagents optimized for performing both sticky and blunt-end ligations. The kit is designed to support up to 150 reactions and includes specific control DNAs and T4 DNA Ligase.
The manual outlines storage requirements for all components at -20°C or lower. It details pre-protocol considerations, such as avoiding high NaCl concentrations, and provides step-by-step protocols for ligating insert DNA into lambda vector arms and plasmid vectors, including methods to verify reaction activity using control ligations.
| Reactions per kit | 150 ligations |
|---|---|
| T4 DNA Ligase activity | 4 U/ul |
| rATP concentration | 10 mM |
| Ligase buffer concentration | 10x |
Most components should be stored at -20°C, though long-term storage for some reagents may require -80°C.
Perform a control ligation using the provided wild-type lambda or pUC18 plasmid control DNA.
Incubate at 12°C for 4-6 hours or at 4°C overnight.