T4 RNA Ligase 2
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.
Key benefits
- 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 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 (blue arrow)was hybridized to an 18-nt chimeric DNA6-RNA4-DNA8 single-stranded oligonucleotide (red arrow) and treated with RNase H resulting in cleavage of the 55-nt ssRNA oligonucleotide into 34-nt (green arrow) & 21-nt (purple arrow) 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.
| Components |
Volume |
T4 RNA Ligase 2, 10 U/μl
in 50% glycerol, 50 mM Tris-HCl, pH 7.5, 0.1 M NaCl, 1 mM
dithiothreitol, 0.1 mM EDTA and 0.1% Triton® X-100. |
100 μl |
10X T4 RNA Ligase 2 Reaction Buffer
0.5 M Tris-HCl, pH 7.5, 20 mM MgCl2. |
500 μl |
|
10 mM ATP |
200 μl |
|
100 mM Dithiothreitol (DTT) |
100 μl |
Manual
SDS
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