T4 RNA Ligase 2
- ATP-dependent RNA ligase that catalyzes phosphodiester bond formation between nucleic acids.
- Highly active on nicked dsRNA.
- Use for nick ligation of double-stranded nucleic acid molecules and ssRNA circularization with a splint molecule.
Product is for research use only (RUO)
Product Description:
T4 RNA Ligase 2 is an ATP-dependent RNA ligase utilizing RNA as the acceptor molecule in nicked double-stranded (ds) nucleic acid molecules using RNA or DNA as the donor molecule and/or as the splinting molecule (nick ligation). The enzyme catalyzes the formation of a phosphodiester bond between a 5'-phosphoryl-terminated nucleic acid donor to a 3'-hydroxyl-terminated nucleic acid acceptor in a template-dependent manner. T4 RNA Ligase 2 is highly active on nicked dsRNA and has diminished activity on single-stranded (ss) RNA substrates.
T4 RNA Ligase 2 can be used for the following applications:
- Nick Ligation of ds nucleic acid molecules with an RNA acceptor strand.
- ssRNA circularization in conjunction with a splint molecule.
- ssRNA end tagging in conjunction with a splint molecule.
Product Performance:
Figure 1. Nick Ligation of various double-stranded substrates by T4 RNA Ligase 2. T4 RNA Ligase 2 utilizes RNA as the acceptor molecule in nicked double-stranded nucleic acid molecules using RNA (blue) or DNA (red) as the donor molecule and/or as the splinting molecule for nick ligation.
Figure 2. A 55-nt ssRNA oligonucleotide (⬅) was hybridized to an 18-nt chimeric DNA6-RNA4-DNA8 single-stranded oligonucleotide (⬅) and treated with RNase H resulting in cleavage of the 55-nt ssRNA oligonucleotide into 34-nt (⬅) & 21-nt (⬅) fragments. The reaction products were purified and treated with T4 RNA
Ligase 2. The 18-nt chimeric DNA6-RNA4-DNA8 single-stranded oligonucleotide acts as a nick ligation splint for rescue of the 55-nt ssRNA oligonucleotide upon ligation.
