Description: T4 DNA ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphoryl and 3'-hydroxyl termini in duplex DNA. It repairs single-strand nicks in duplex DNA and will join both blunt-ended and cohesive-ended restriction fragments of duplex DNA or RNA. The enzyme requires ATP as cofactor.
Storage conditions: 25 mM Tris-HCl, pH 7.6, 100 mM NaCl, 1 mM DTT, 0.5 mM EDTA and 50% (v/v) glycerol. Store at -20 °C in a non-frost free freezer.
Concentration: 5 U/µl
10× reaction buffer: 660 mM Tris-HCl, pH 7.6, 66 mM MgCl2, 100 mM DTT, 660 µM ATP. Vortex the reaction buffer solution thoroughly after thawing and prior to use. Repeated freeze-thaw cycles will affect the stability of ATP. We recommend making up 10-20 µl aliquots of the buffer and storage at -20 °C.