T4 RNA Ligase I, 1000 U

Product Information

Product Description/Introduction

T4RNA ligase I, derived from the RNA ligase I gene of phage T4, is reexpressed by Escherichia coli. It is an ATP-dependent enzyme that catalyzes the formation of intermolecular or intramolecular phosphodiester bonds between the 5' phosphate terminal and the 3' hydroxyl terminal of single stranded RNA, single stranded DNA, or single nucleotides. T4RNAligase I is also considered a single stranded RNA (ssRNA) ligase because it is the most efficient for linking single-stranded RNA, followed by single-stranded DNA to RNA (when DNA provides a 5 'phosphate group and RNA provides a 3' hydroxyl group), and the least efficient for linking single-stranded DNA.

Uses:ssRNA intermolecular junction; Intermolecular linkage between ssDNA and ssRNA; ssRNA end markers, etc.

Source: Derived from phage T4 recombinant expression by Escherichia coli.

Enzyme storage buffer:10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4.

10×RNA ligaseBuffer：500 mM Tris-HCl, 100 mM MgCl₂, 10 mM DTT, pH 7.5.

Storage and Shipping Conditions

Wet ice transport; Store at -20 for 12 months.

Product Content

Assay Protocol / Procedures

1. For cyclization linking reactions of single-stranded RNA, reference reaction system:

Note: PEG8000 (final concentration 15%-25%) can be added to the reaction system to significantly improve enzyme activity without affecting the specificity of the reaction. All reagents and operations should avoid RNase contamination; The amount of RNA substrate in the reference reaction system has been relatively large, and the amount of ssRNA can be greatly reduced when the sample is relatively small.

2. Incubate the above reaction at 37 for 30min (in order to make the connection more adequate, the reaction time can be appropriately extended). The reaction conditions can be adjusted according to the actual situation. If the effect is not satisfactory, incubation at 25 for 2h or 16 for 16h can be tried.

3. After the reaction was completed, the reaction system was incubated at 65 for 15min to terminate the reaction. Perform subsequent operations as required.

Note

1. Involving RNA operation, it is necessary to strictly follow the specifications of RNA operation to avoid RNase contamination, and related reagents and consumables need to be treated by DEPC to remove RNase or ensure that it is RNase free.

2. RNase inhibitors can be used to prevent degradation of RNA during the reaction. An RNase inhibitor (G3414) with a final concentration of 1U/μL can be added.

3. Enzyme products should be placed in an ice box or ice bath when used, and should be stored at -20 immediately after use, it is recommended to store in separate packages.

4. For your safty and health,please wear safety glasses, gloves, or protective clothing.

For Research Use Only!

https://www.servicebio.com/goodsdetail?id=72622