Thermostable 5´ AppDNA/RNA Ligase
Thermostable 5´ App DNA/RNA Ligase ligates a 5´ pre-adenylated 3´-blocked DNA or RNA sequence tag to the 3´-OH of RNA and ssDNA.
- Isolated from a recombinant source
- Supplied with 10X reaction buffer
- Tested for the absence of DNases and RNases
- Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart
Avoiding RNase Contamination
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 SourceThermostable 5´ App DNA/RNA Ligase is expressed as His-tag fusion in E. coli.
The following reagents are supplied with this product:
Store at (°C) Concentration MnCl2 -20 10 X NEBuffer™ 1 -20 10 X
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