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

Descripción general

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

Sinónimos

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

Especificaciones y pureza

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

Estabilidad y almacenamiento

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

Condiciones de almacenamiento de almacenamiento

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

Enviado en

Ice chest + Ice pads

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Grado

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