T4 RNA ligase

This product is an ATP-dependent T4 RNA ligase (T4 RNA Ligase I) recombinantly expressed by E coli. This enzyme catalyzes the formation of phosphodiester bonds between oligonucleotides, single-stranded RNA and DNA intermolecular/ intra-molecular 5′-PO4 and 3′-OH.

Product component

Storage condition

Store at -25～-15°C (avoid repeated freezing and thawing)

Storage buffer

50 mM KCl, 10 mM Tris-HCl, 1 mM DTT, 0.1mM EDTA, 50 % Glycerol, (pH 7.4 @ 25°C)

Unit definition

One unit is defined as the amount of enzyme required to convert 1 nanomole of 5´-[³²P] rA16 into a phosphatase-resistant form in 30 minutes at 37°C.

Quality control

Purity ≥ 95%, host DNA residue ≤100 pg/mg, host protein residue ≤ 50 ppm, Endotoxin residue ≤10 EU/mg, no RNase, Endonuclease, Exonuclease, Protease residue, Sterile, Mycoplasma free.

Reaction system and conditions

single-stranded RNA intermolecular ligation Prepare the reaction system according to the following table:

2. 1. After mixing, Incubate at 37°C for 30 minutes;
3. 2. Incubate at 65°C for 15 min to terminate the reaction.

1. 1. For your safety and health, please wear lab coat and disposable gloves.

2. 2. RNase contamination should be avoided during the reaction process, and related reagents and consumables should be treated with DEPC to remove RNase.

3. 3. Incubate at 25°C for 2h or 16h at 16°C during the reaction. The ligation reaction time can be appropriately extended to make the connection more sufficient.

4. 4. The amount of ssRNA can be optimized according to the sample volume.