T4 RNA Ligase II, also known as T4 Rnl2 (gp24.1), catalyses the formation of an internucleotide phosphodiester bond between an oligonucleotide donor molecule with a 5′-terminal phosphate and an oligonucleotide acceptor molecule with a 3′-terminal hydroxyl. 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 nucleic acids. T4 RNA ligase II is expressed and purified from E.coli.

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 & Storage Condition
- T4 RNA Ligase II : 10 unit/㎕, Store at -20°C.
- 10x T4 RNA Ligase II Reaction Buffer : Store at 4°C.

Reaction Condition
- T4 RNA Ligae II in 1X T4 RNA RNA ligase II Reaction Buffer. Incubate at 37°C.

10x Reaction Buffer
- 500 mM Tris-HCl (pH 7.5), 20 mM MgCl₂, 10 mM DTT, 4 mM ATP.

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.

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.

QC Tests

- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.