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FAQs for T4 RNA Ligase 1 (ssRNA Ligase) Protocols for T4 RNA Ligase 1 (ssRNA Ligase) 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 Reaction Buffer T4 RNA Ligase 2 (dsRNA Ligase)

Home > Products > RNAi and RNA Enzymes > RNA Ligases > T4 RNA Ligase 1 (ssRNA Ligase) T4 RNA Ligase 1 (ssRNA Ligase) Catalog # Size Concentration Price Qty   M0204L 5,000 units 20,000 units/ml $212.00 M0204S 1,000 units 20,000 units/ml $53.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Isolated from a recombinant source Ligation of a single-stranded RNA and DNA Labeling of the 3' ends of RNA with 5'-[32P]pCp Synthesis of Single-stranded Oligonucleotides Supplied with 10X Reaction Buffer Description: T4 RNA Ligase 1 catalyzes the ligation of a 5' phosphoryl-terminated nucleic acid donor to a 3' hydroxyl-terminated nucleic acid acceptor through the formation of a 3'→5' phosphodiester bond, with hydrolysis of ATP to AMP and PPi. Substrates include single-stranded RNA and DNA as well as dinucleoside pyrophosphates (1). Source: Purified from E. coli strain ER2497 containing the plasmid pRF-E35 [constructed at New England Biolabs, Inc. after the method of K. N. Rand and M. J. Gait (2)]. Applications: Labeling of 3'-termini of RNA with 5'-[32P] pCp (3) Inter- and intramolecular joining of RNA and DNA molecules (4,5) Synthesis of single-stranded oligodeoxyribo-nucleotides (6) Incorporation of unnatural amino acids into proteins (7) Reagents Supplied: T4 RNA Ligase 1 Reaction Buffer (10X) Enzyme Properties Heat Inactivation: 65°C for 15 minutes Heat inactivation by boiling for 2 minutes. Specific Activity: 12,000 units/mg Reaction & Storage Conditions Reaction Conditions: 1X T4 RNA Ligase 1 Reaction Buffer Incubate at 37°C. 1X T4 RNA Ligase 1 Reaction Buffer: 50 mM Tris-HCl 10 mM MgCl2 1 mM ATP 10 mM Dithiothreitol pH 7.8 @ 25°C Unit Definition: One unit is defined as the amount of enzyme required to convert 1 nmol of 5'-phosphoryl termini in 5'-[32P]rA20 to a phosphatase-resistant form in a total reaction volume of 50 μl in 30 minutes at 37°C (1). Unit Assay Conditions: 1X T4 RNA Ligase 1 Reaction Buffer and RNA (10 µM 5´-[32P]rA20, 10 µM in 5´ termini). After incubation at 37°C for 15 minutes, the reaction is terminated by boiling for 2 minutes, and the bacterial alkaline phosphatase-resistant 5´ phosphoryl termini are determined as described (8). Concentration: 20,000 units/ml Storage Conditions: 10 mM Tris-HCl 50 mM KCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Storage Temperature: -20°C Notes Usage notes: Addition of DMSO to 10% (v/v) is required for pCp ligation (3). FAQs Can T4 RNA Ligase 1 be used to end label single stranded DNA or RNA? Will T4 RNA Ligase 1 ligate double stranded DNA? Will T4 RNA Ligase 1 ligate single stranded DNA? What is the most common cause of ligation failure when using T4 RNA Ligase 1? Will the ligation reaction using T4 RNA Ligase 1 produce circles or duplex products? Can T4 RNA Ligase 1 be used to add tails to linear duplex DNA? Can T4 RNA Ligase 1 be used to make single stranded RNA/DNA hybrids? Can T4 RNA Ligase 1 ligate triphosphates? What is the specific activity of T4 RNA Ligase 1? Will PEG improve ligation with T4 RNA Ligase 1? Can a dideoxy be used to block ligation using T4 RNA Ligase 1? Can T4 RNA Ligase 1 be used to incorporate unnatural amino acids into proteins? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Quality Assurance Statement: Each lot is tested for contaminating single-stranded DNA exonuclease, endonuclease, ribonuclease and phosphatase activities. Exonuclease Activity: Incubation of a 50 μl reaction containing 50 units of T4 RNA Ligase 1 (ssRNA Ligase) with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity. Endonuclease Activity: Incubation of a 50 μl reaction containing 50 units of T4 RNA Ligase 1 (ssRNA Ligase) with 1 μg of ΦX174 RF I DNA for 4 hours at 37ºC resulted in < 10% conversion to RFII as determined by agarose gel electrophoresis. Ribonuclease Activity: Incubation of 50 units of T4 RNA Ligase 1 with 3 μg of ssRNA ladder (NEB #N0362) in 50 μl T4 RNA Ligase 1 Reaction Buffer for 3 hours at 37°C resulted in no detectable degradation of the RNA as determined by agarose gel electrophoresis. References England, T., Gumport, R. and Uhlenbeck, O. (1977) Proc. Natl. Acad. Sci. USA, 74, 4839-4842. Rand, K.N. and Gait, M.J. (1984) EMBO J., 3, 397-402. England, T. and Uhlenbeck, O. (1978) Nature, 275, 560-562. Romaniuk, P. and Uhlenbeck, O. (1983) R. Wu, L. Grossman and K. Moldave (Eds.), Methods Enzymol., 100, pp. 52-56. New York: Academic Press. Moore, M.J. and Sharp, P.A. (1992) Science, 256, 992-997. Tessier, D.C., Brousseau, R., and Vernet, T. (1986) Anal. Biochem., 158, 171-178. Noren, C.J. et al. (1989) Science, 244, 182-188. Sugino, A. et al. (1977) J. Biol. Chem., 252, 1732-1783. Reagents Sold Separately T4 RNA Ligase 1 Reaction Buffer Companion Products T4 RNA Ligase 2 (dsRNA Ligase)