T4 RNA Ligase 2 (dsRNA Ligase)

Description: 
T4 RNA Ligase 2, also known as T4 Rnl2 (gp24.1), catalyzes both inter- and intramolecular formation of phosphodiester bonds between 5'-phosphate and 3'-hydroxyl termini of oligonucleotides, single-stranded RNA and DNA. Unlike T4 RNA Ligase 1, T4 RNA Ligase 2 is much more active on joining nicks on double stranded RNA than on joining the ends of single-stranded RNA. The enzyme requires an adjacent 5'-phosphate and 3'-OH for ligation. The enzyme can also ligate the 3'-OH of RNA to the 5'-phosphate of DNA in a double stranded structure.

Source:
An E. coli strain that carries the T4 RNA Ligase 2 gene (I. Schildkraut).

Application:
- Ligates a nick in dsRNA
- Ligates the 3'-OH of RNA to the 5'-phosphate of DNA in a double stranded structure

Reagents Supplied: 
T4 Rnl2 Reaction Buffer

Reaction Conditions: 
1x T4 Rnl2 Reaction Buffer
Incubate at 37°C

10x T4 RNA Ligase 2 Reaction Buffer:
500 mM Tris-HCl
20 mM MgCl₂
10 mM DTT
4 mM ATP
pH 7.5 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to ligate 0.4 µg of an equimolar mix of a 23-mer and 17-mer RNAs in a total reaction volume of 20 µl in 30 minutes at 37°C.

Unit Assay Conditions:
1x T4 Rnl2 Reaction Buffer and 0.4 µg of an equimolar mix of the 23-mer and 17-mer RNAs. After incubation at 37°C for 30 minutes, the ligated product is detected on a 15% polyacrylamide gel.

Specific Activity: >40,000U/mg
 

Storage Conditions:
10 mM Tris-HCl
50 mM KCl
35 mM (NH₄)₂SO₄
0.1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Recommended Storage Condition: -20°C