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Thermostable 5´ App DNA/RNA Ligase is a point mutant of catalytic lysine of RNA ligase from Methanobacterium thermoautotrophicum (1). 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 (1). The optimal temperature for ligation reaction is 60–65°C (2). 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.- This product is related to the following categories:
- RNA Ligation,
- DNA Ligases Products
- This product can be used in the following applications:
- Non-Cloning Ligation
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