T4 RNA Ligase 1 - BioReagent, PCR Reagent, for DNA synthesis, Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications, 30 U/μl

Cat. No.: T750872

GRADE & PURITY Suitable for molecular biology, BioReagent, PCR Reagent, for DNA synthesis, EnzymoPure™, for DNA and RNA applications, 30 U/μl

Synonyms

T4 RNA Ligase 1 | T4 RNA Ligase 1 (ssRNA Ligase)

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Application

PCR

Overview

Storage Buffer： 10mM Tris-HCl, 50mM KCl, 0.1mM EDTA, 1mM DTT, 50% Glycerol, pH 7.5  

T750872	Component	600U	3000 U	Storage
T750872A	T4 RNA Ligase 1	20 μL	100 μL	-20°C. Avoid freeze/thaw cycle
T750872B	10×T4 RNA Ligase 1 Buffer	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872C	50% PEG 8000	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872D	ATP	10 μL	50 μL	-20°C. Avoid freeze/thaw cycle

T4 RNA Ligase 1 is an ATP-dependent single-stranded nucleic acid molecule (DNA or RNA) ligase that catalyzes the 5' phosphate group of the donor and the 3' hydroxyl group of the ligand to form 3'→5' phosphodiester bonds and accompanies ATP hydrolysis to AMP and PPi. The substrates of this enzyme are ssRNA, ssDNA and dinucleoside pyrophosphate.

Enzyme Activity Definition
At 37℃, the amount of enzyme required to convert 1 nmol of 5 '-[32P] rA16 into phosphate insoluble matter within 30 min is defined as 1 unit (U).
Protocol

1. Cyclization of RNA
(1) Prepare the mixed solution as shown in the following table:

Component	Volume
ssRNA	200 ng-1 μg
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	20-50 μM
50% PEG 8000	10% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2) Incubation at 25 °C for 2h or 16°C for 16h.
(3) The reaction was terminated by boiling at 65 °C for 15 min or 2min.
2. Linking reaction between RNA or DNA oligonucleotides and ssRNA
(1) Prepare the mixed solution as shown in the following table:

Component	Volume
ssRNA	1-20 pmol
RNA or DNA oligonucleotides	5-40 pmol
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	2 μl
50% PEG 8000	15% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2) Incubation at 25°C for 2h or 16°C for 16h.
(3) The reaction was terminated at 65°C15 min or by boiling for 2min.
Cautions
This product is for scientific research purposes only and shall not be used for other purposes.

Specifications

Synonyms

T4 RNA Ligase 1 | T4 RNA Ligase 1 (ssRNA Ligase)

Specifications & Purity

BioReagent, PCR Reagent, for DNA synthesis, Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications, 30 U/μl

Stability And Storage

Store at -20℃ long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Storage

Store at -20°C, Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent, for DNA and RNA applications, for DNA synthesis, PCR Reagent, Suitable for molecular biology, EnzymoPure™