Thermo Scientific™ T4 DNA Ligase
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.
Manufacturer: Thermo Scientific™ EL0016
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA. The enzyme repairs single-strand nicks in duplex DNA, RNA, or DNA/RNA hybrids. It also joins DNA fragments with either cohesive or blunt termini, but has no activity on single-stranded nucleic acids.
The T4 DNA Ligase requires ATP as a cofactor.
- Active in Themo Scientific restriction enzyme, PCR, and RT buffers (when supplemented with ATP)
- Fast – sticky-end ligation is completed in 10 minutes at room temperature
- Supplied with PEG solution for efficient blunt-end ligation
- Cloning of restriction enzyme generated DNA fragments
- Cloning of PCR products
- Joining of double-stranded oligonucleotide linkers or adaptors to DNA
- Site-directed mutagenesis
- Amplified fragment length polymorphism (AFLP)
- Ligase-mediated RNA detection (see Reference 3)
- Nick repair in duplex DNA, RNA or DNA/RNA hybrids
- Self-circularization of linear DNA.
- Binding of T4 DNA Ligase to DNA may result in a band shift in agarose gels. To avoid this, incubate samples with 6X DNA Loading Dye & SDS Solution at 70°C for 5 min or 65°C for 10 minutes and chill on ice prior to electrophoresis.
- The volume of the ligation reaction mixture should not exceed 10% of the competent cell volume in the transformation process.
- Prior to electro-transformation, remove T4 DNA Ligase from the ligation mixture by spin column or chloroform extraction. The extracted DNA can be further precipitated with ethanol.
|E.coli cells with a cloned gene 30 of bacteriophage T4|
|55.3 kDa monomer|
|T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration exceeds 200mM.|
|LC; 2 x 500U|