T4 RNA Ligase 2, Truncated

T4 RNA Ligase 2 truncated catalyzes phosphodiester bond formation between a pre-adenylated 5′ phosphate (DNA or RNA) and the 3′ hydroxyl of RNA. The truncated enzyme contains the first 249 amino acids which makes the enzyme require a pre-adenylated 5′ terminal donor and eliminates the need for ATP. Because T4 RNA ligase 2 truncated cannot use the 5′ phosphate of RNA or DNA as a donor in the ligation reaction, it is useful for certain applications such as linker ligations with pre-adenylated 5′ DNA to 3′ hydroxyl RNA. The desired specific ligation products are enhanced dramatically over unwanted background ligation products, making the truncated enzyme superior to the full-length enzyme for this use.

Suitable for:

• NGS RNA library construction
• Linker ligations with pre-adenylated 5′ DNA to 3′ hydroxyl RNA

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′ pre-adenylated end of a 18-mer DNA when both 17-mers are annealed to a complementary 35-mer DNA strand in 20 μL 1X reaction buffer following a 30 minute incubation at 37° C.

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

Supplied with:KEM0023B (10X T4 RNA Ligase 2, Truncated Buffer)

Unit size: 500 U