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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.
Source of ProteinPurified from a strain of E. coli that expresses the recombinant truncated T4 RNA Ligase 2 gene.
Supplied in10 mM Tris-HCl100 mM NaCl0.1 mM DTT0.1 mM EDTA50% glycerolpH 7.5 @ 25°C
Supplied With:B6070 10X T4 RNA Ligase 2, Truncated Buffer
10X T4 RNA Ligase 2, Truncated Buffer (B6070)500 mM Tris-HCl100 mM MgCl250 mM DTTpH 7.6 @ 25°C
Unit DefinitionOne 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.
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