T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration exceeds 200mM.
E.coli cells with a cloned gene 30 of bacteriophage T4
55.3 kDa monomer
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
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.