T4 RNA Ligase 2
T4 RNA Ligase 2
Source:
Recombinant
T4 RNA Ligase 2, also known as T4 Rnl-2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity. T4 RNA Ligase 2 is much more active than T4 RNA Ligase 1 (PN 21245) in 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. T4 RNA Ligase 2 can also ligate the 3’ OH of RNA to the 5’ phosphate of DNA in a double stranded structure.
Purity:
Greater than 90% pure as determined by SDS-PAGE.
Storage buffer:
10 mM Tris-HCl, pH 7.5, 50 mM KCl, 35 mM (NH4)2SO4, 0.1 mM DTT, 0.1 mM EDTA, 50% glycerol
Assay conditions:
The reaction mixture contains 0.4 µg of an equimolar mix of a 23-mer and 17-mer RNAs, T4 RNA Ligase 2 in 1X T4 RNA Ligase 2 Reaction Buffer at 37°C in 30 minutes (20 µl reaction volume).
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 RNA at 37°C in 30 minutes.
Concentration:
10 units/µl
Functionally tested 10X T4 RNA Ligase 2 Reaction Buffer (1 ml included):
500 mM Tris-HCl, pH 7.5, 20 mM MgCl2, 10 mM DTT, and 4 mM ATP.
Applications:
• Ideal for ligating nicks in dsRNA
• Ligating the 3’ OH of RNA to the 5’ phosphate of DNA in a double-stranded structure
• Suitable for ligating 3’ OH or RNA to 5’ phosphate of DNA in a DNA/RNA hybrid
Storage:
Shipped on dry ice. Store at -20°C.