T4 DNA Ligase CAS 9015-85-4

English name：T4 DNA Ligase

Grade：BR

Source: This T4 DNA ligase is expressed by E. coli, and the source of the expressed gene is T4 phage.

Activity definition：One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min at 16℃ in 20 μl of the assay mixture containing 50 mM Tris, pH 7.5, 10 mM MgCl2, 10 mM DTT, 1 mM ATP, 25 μg/ml BSA and a 5'-DNA termini concentration of 0.12 μM (300 μg/ml)。

Enzyme storage solution：20 mM Tris, pH 7.5, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA and 50% (v/v) glycerol。

10X Ligation Buffer：400 mM Tris, pH 7.8, 100 mM MgCl2, 100 mM DTT, 5 mM ATP。

Inactivation or inhibition: incubation at 65 ℃ for 10 minutes can inactivate T4 DNA ligase; T4 DNA ligase was strongly inhibited when the concentration of NaCl or KCl was greater than 200 mM.

Physicochemical properties: liquid. The enzyme catalyzes the formation of phosphate diester bond between the adjacent 5 'phosphate group and the 3' - hydroxyl end in double stranded DNA or RNA. It can also repair the single strand incision in double stranded DNA, RNA or DNA / RNA complex to connect the sticky and flat end of DNA, but the enzyme has no enzymatic activity for single stranded nucleic acid. The enzyme requires the cofactor ATP.

Purpose: Biochemical research.Sticky end or flat end double stranded DNA connection; Double stranded oligonucleotide linker is connected with double stranded DNA; Repair of gaps in double stranded DNA, RNA or DNA-RNA complex; Ligase mediated RNA detection; Site specific mutation；Amplified fragment length polymorphism.