T4 RNA Ligase 2 (dsRNA Ligase)

Description: 
T4 RNA Ligase 2, also known as T4 Rnl2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity. Unlike T4 RNA Ligase 1, T4 RNA Ligase 2 is much more active 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).

Specific Activity:
40,000U/mg

• Ligate a nick in dsRNA

• Ligate 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.

1X T4 Rnl2 Reaction Buffer:
50 mM Tris-HCl
2 mM MgCl2
1 mM DTT
400 µM 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.

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 Conditions:
 -20°C