M0319L, Thermostable 5'AppDNA/RNA Ligase - 50 reactions

Thermostable 5´ App DNA/RNA Ligase is a point mutant of catalytic lysine of RNA ligase from Methanobacterium thermoautotrophicum. This enzyme is ATP independent. It requires a 5´ pre-adenylated linker for ligation to the 3´-OH end of either RNA or single stranded DNA (ssDNA). The enzyme is also active in ligation of RNA with 2´-O-methylated 3´ end to 5´-adenylated linkers. The optimal temperature for ligation reaction is 60–65°C. The mutant ligase is unable to adenylate the 5´-phosphate of RNA or ssDNA, which reduces the formation of undesired ligation products (concatemers and circles).

The ability of the ligase to function at 65°C might reduce the constraints of RNA secondary structure in RNA ligation experiments.

Product Source

Thermostable 5´ App DNA/RNA Ligase is expressed as His-tag fusion in E. coli.