T4 RNA Ligase 2

T4 RNA Ligase 2 catalyzes phosphodiester bond formation between a 5′ phosphate and 3′ hydroxyl of RNA. The preferred substrate is nicked double-stranded RNA but single-stranded RNA can also serve as a substrate. Ligation of single-stranded RNA substrates generates either intramolecular or intermolecular products. Besides nicked double-stranded RNA substrates, other nicked nucleic acids hybrids can be sealed. The strand containing the 5′ phosphate can either be DNA or RNA. The non-ligated strand of the duplex can be either RNA or DNA. T4 RNA ligase 2 requires ATP for activity unless the substrate is preadenylated on the 5′ end. A truncated version of T4 RNA ligase 2 is a better enzyme for preadenylated substrates because it generates less side-reaction ligation products than the full length enzyme.

Suitable for:

• Nick ligation of dsRNA
• Ligation of 3′OH RNA to 5′ phosphate of DNA (in ds format)

Supplied in 10 mM Tris-HCl, 100 mM NaCl, 0.1 mM DTT, 0.1 mM EDTA and 50% glycerol at pH 7.5 @ 25° C.

One unit is defined as the amount of enzyme required to ligate 50% of 0.4 μg of an equimolar mix of a single-stranded 5′ FAM-labeled 17-mer RNA to the 5′ phosphorylated end of a 18-mer DNA when both strands are annealed to a complementary 35-mer DNA strand in 20 μL at 37° C for 30 minutes.

Source of protein: Purified from a strain of E. coli that expresses the recombinant T4 RNA Ligase 2 gene.

Supplied with:KEM0049B (10X Ligation Buffer)

Unit size: 4,500 U