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FAQs for RNAi and RNA Enzymes Technical Reference for RNAi and RNA Enzymes Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE T4 RNA Ligase 1 (ssRNA Ligase)

Home > Products > RNAi and RNA Enzymes > RNA Ligases > T4 RNA Ligase 2 (dsRNA Ligase) T4 RNA Ligase 2 (dsRNA Ligase) Catalog # Size Concentration Price Qty   M0239L 750 units 10,000 units/ml $272.00 M0239S 150 units 10,000 units/ml $68.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Best choice for ligating nicks in dsRNA Suitable for ligating 3' OH or RNA to 5' phosphate of DNA in a DNA/RNA hybrid Isolated from a recombinant source Supplied with 10X Reaction Buffer Description: T4 RNA Ligase 2, also known as T4 Rnl-2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity (1-3). Unlike T4 RNA Ligase 1 (NEB #M0204), 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 (4). Source: An E. coli strain that carries the T4 RNA Ligase 2 gene. Applications: 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 RNA Ligase 2 Reaction Buffer Enzyme Properties Heat Inactivation: No Specific Activity: 40,000 units/mg Reaction & Storage Conditions Reaction Conditions: 1X T4 RNA Ligase 2 Reaction Buffer Incubate at 37°C. 1X T4 RNA Ligase 2 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 RNA Ligase 2 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. Concentration: 10,000 units/ml 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 Storage Temperature: -20°C Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Ribonuclease Activity: Incubation of 90 units of T4 RNA Ligase 2 with 3 µg of ssRNA ladder (NEB #N0362) in 50 µl T4 RNA Ligase 2 Reaction Buffer for 3 hours at 37°C resulted in no detectable degradation of the RNA as determined by agarose gel electrophoresis. Phosphatase Activity: Incubation of 150 units of T4 RNA Ligase 2 with 1 µg p-nitrophenyl phosphate (pNPP) in 50 µl T4 RNA Ligase 2 Reaction Buffer for 16 hours at 37°C released less than 0.05 µmol inorganic phosphate. DNA Endonuclease Activity: Incubation of 150 units of T4 RNA Ligase 2 with 1 μg of ΦX174 RF I DNA in 50 μl for 4 hours at 37ºC resulted in no detectable degradation of DNA as determined by agarose gel electrophoresis. DNA Exonuclease Activity: Incubation of 150 units of T4 RNA Ligase 2 with 1 μg of a mixture of single and double-stranded sonicated 3H DNA (105 cpm/μg) in 50 μl T4 RNA Ligase 2 Reaction Buffer for 4 hours at 37ºC released 0.1% of the activity. References Ho, C.K. and Shuman, S. (2002) Proc. Natl. Acad. Sci. USA, 99, 12709-12714. Ho, C.K. et al. (2004) Structure, 12, 327-339. Nandakumar, J. et al. (2004) J. Biol. Chem., 279, 31337-31347. Nandakuman, J. and Shuman, S. (2004) Mol. Cell, 16, 211-221. Companion Products T4 RNA Ligase 1 (ssRNA Ligase)