Thermo Scientific™ Rapid DNA Ligation Kit

The Thermo Scientific™ Rapid DNA Ligation Kit enables rapid sticky-end or blunt-end DNA ligation in just 5 minutes at room temperature. This kit contains T4 DNA ligase and a specially formulated 5X rapid ligation buffer, optimized to provide fast and efficient DNA ligation. The ligation efficiency achieved with this kit is comparable to a traditional 1-hour ligation with T4 DNA ligase. The ligation reaction mixture can be directly used for bacterial transformation, including with the TransformAid Bacterial Transformation Kit, or with other conventional transformation methods.

Key Features & Benefits:

1. Fast Ligation:
   • The kit allows for the ligation of both sticky-end and blunt-end DNA in just 5 minutes at room temperature, significantly reducing the time required for DNA ligation compared to traditional methods.
2. High Ligation Efficiency:
   • Provides ligation efficiency comparable to standard 1-hour ligation reactions using T4 DNA ligase, making it a time-saving solution for molecular cloning.
3. Convenient for Transformation:
   • The ligation reaction mixture can be used directly for bacterial transformation, saving time and eliminating the need for additional purification steps. It's compatible with the TransformAid Bacterial Transformation Kit or other transformation procedures.
4. Optimized Buffer System:
   • The 5X Rapid Ligation Buffer is specially formulated to optimize ligation efficiency, ensuring fast and reliable results.
5. Versatile Applications:
   • Ideal for routine cloning experiments, blunt-end cloning, library construction, and TA cloning, this kit is perfect for a wide range of molecular biology applications.

Applications:

• Routine Cloning Experiments: Ideal for constructing recombinant plasmids and vectors.
• Blunt-End Cloning: Enables ligation of blunt-ended DNA fragments, useful for cloning when restriction enzymes create blunt ends.
• Library Construction: Useful in constructing DNA libraries, ensuring efficient ligation of DNA fragments into vectors.
• TA Cloning: Perfect for TA cloning applications, especially when using Taq polymerase to create overhanging A-tailed DNA.

Includes:

• T4 DNA Ligase: The enzyme required for catalyzing the formation of the phosphodiester bond between DNA fragments.
• 5X Rapid Ligation Buffer: The optimized buffer for fast and efficient ligation.
• Nuclease-free Water: To dilute the reaction components as needed.
• Detailed Protocol: Provides step-by-step instructions to guide the user through the process.

Important Notes:

• Inactivation of T4 DNA Ligase: Before electroporation, T4 DNA ligase must be inactivated by chloroform extraction or spin column purification, such as using the Thermo Scientific GeneJET PCR Purification Kit (K0701).

Specifications:

Storage and Handling:

• Storage: Store the kit components at -20°C for long-term storage to maintain stability. The 5X Rapid Ligation Buffer and T4 DNA Ligase should be stored at -20°C, while the nuclease-free water can be stored at ambient temperature.

The Thermo Scientific™ Rapid DNA Ligation Kit is a powerful tool for fast and efficient DNA ligation in a variety of molecular biology applications, including cloning, TA cloning, and library construction. Its ability to perform ligation in just 5 minutes at room temperature makes it an ideal solution for researchers needing to streamline their cloning and transformation processes, saving valuable time while maintaining high efficiency.