T4 RNA Ligase 1,UA BIOSCIENCE_specification/price/image

Product Detail
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Product Specification

Synonyms	T4 RNA Ligase, T4 Rnl1
Expression System	E.coli
Molecular Weight	69kDa (Reducing)
Purity	＞95% by SDS-PAGE
Tag	His Tag
Storage Buffer	10 mM Tris-HCl、50 mM KCl、1 mM DTT、0.1 mM EDTA、50% Glycerol pH 7.4 @ 25°C
Stability & Storage	Store at -25 ~ -15℃ for 2 years
Reference	[1] England, Thomas E , and O. C. Uhlenbeck . "3\|[prime]\|-Terminal labelling of RNA with T4 RNA ligase." Nature 275.5680(1978):560-1. [2] Tessier, Daniel C. , R. Brousseau , and T. Vernet . "Ligation of single-stranded oligodeoxyribonucleotides by T4 RNA ligase." Analytical Biochemistry 158.1(1986):171-178.

Background

T4 RNA Ligase 1 is an ATP-dependent enzyme that catalyzes the formation of phosphodiester bonds between the 5'-P and 3'-OH ends of single-stranded RNA, single-stranded DNA, or single nucleotides, either intermolecular or intramolecular. It is suitable for the connection of single-stranded RNA molecules and the 5' end junction constructed by miRNA libraries in NGS. Labeled with RNA 3 'terminal; Cyclization of oligonucleotides; tRNA modification; In 5'-RACE (Rapid Amplification of cDNA Ends) assay, it is used for oligonucleotides to bind to single-stranded cDNA; and the introduction of unnatural amino acids into proteins.

Components

Name	Storage Temperature	quantity	concentration
T4 RNA Ligase 1	-20 ℃	100 ul or 500 µl	10 U/µl
10*Reaction Buffer	-20 ℃	1 ml	10 X
Adenosine 5'-Triphosphate (ATP)	-20 ℃	1 ml	10 mM
PEG8000 (RNase free)	-20 ℃	1 ml	50%

Protocol

1. Set up a 20 μl reaction as follows:

Reagent	Volume	Final Concentration
10 * Reaction Buffer	2 ul	1 X
Single-stranded RNA with 5´P and 3´OH ends (200 ng-1 µg)	1 ul	10-50 ng/ul
50% PEG8000	4 ul	10%
1mM ATP	1 ul	50 µM
40 μ/ul Rnase inhibitor	0.5ul	1 U/µl
10 U/µl T4 RNA Ligase 1	1ul	0.5 U/µl
DEPC-treated Water	Up to 20ul	-

2. Incubate at 37°C for 1-2 hours.
For longer oligos, overnight incubation at 16°C for 16h may improve yield.
3. Incubate at 65°C for 15min to terminate the reaction.

Guidelines

Please avoid repeated freeze-thaw cycles.

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.

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.