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T4 RNA Ligase
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Molecular Biological Enzymes
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Product description
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Product Description
T4 RNA Ligase 1 is an ATP-dependent enzyme that can catalyze the formation of phosphodiester bonds between ssRNA, ssDNA, or single nucleotide molecules or between 5´ phosphate ends and 3´ hydroxyl ends. During this process, ATP is hydrolyzed to AMP and PPi. This product is derived from the E.coli strain carrying the T4 RNA ligase 1 gene.
Source: Recombinant Escherichia coli.
Specification: 10 U/µL.
application
1. RNA 3´-end labeling;
2. Intramolecular or intermolecular connection between ssRNA and ssDNA;
3. Synthesis of single-stranded oligodeoxynucleotides;
4. RNA circularization.
Transportation and storage methods
Ice pack transportation. Store the product at -20°C.
Activity definition
At 37°C, the amount of enzyme required to convert 1nmol of 5´-[32P] rA16 into phosphate insoluble matter within 30 minutes is defined as 1 activity unit (U).
Quality Inspection
No endo or exonuclease, ribonuclease, phosphatase contamination.
Operation Manual
RNA circularization:
1. 20μL reaction system:
T4 RNA Ligase 1 is an ATP-dependent enzyme that can catalyze the formation of phosphodiester bonds between ssRNA, ssDNA, or single nucleotide molecules or between 5´ phosphate ends and 3´ hydroxyl ends. During this process, ATP is hydrolyzed to AMP and PPi. This product is derived from the E.coli strain carrying the T4 RNA ligase 1 gene.
Source: Recombinant Escherichia coli.
Specification: 10 U/µL.
application
1. RNA 3´-end labeling;
2. Intramolecular or intermolecular connection between ssRNA and ssDNA;
3. Synthesis of single-stranded oligodeoxynucleotides;
4. RNA circularization.
Transportation and storage methods
Ice pack transportation. Store the product at -20°C.
Activity definition
At 37°C, the amount of enzyme required to convert 1nmol of 5´-[32P] rA16 into phosphate insoluble matter within 30 minutes is defined as 1 activity unit (U).
Quality Inspection
No endo or exonuclease, ribonuclease, phosphatase contamination.
Operation Manual
RNA circularization:
1. 20μL reaction system:
| Reagent | Sample volume |
| 10× T4 RNA Ligase Reaction Buffer | 2μL |
| 50% PEG 8000 | 4μL |
| ATP (10 mM) | 0.04-0.1μL |
| RNase inhibitor (40 U/μL) | 0.5μL |
| RNA(1 mM) | 0.2μL |
| T4 RNA Ligase (10 U/μL) | 1μL |
| ddH2O | Up to 20μL |
2. Reaction procedures
1) Incubation: 25°C, 1-2 h. For longer oligonucleotides, overnight at 16°C can increase yield.
2) Stop the reaction: 95°C, 2 min or 65°C, 15 min.
Precautions
1. For your safety and health, please wear lab coats and disposable gloves for operation.
1) Incubation: 25°C, 1-2 h. For longer oligonucleotides, overnight at 16°C can increase yield.
2) Stop the reaction: 95°C, 2 min or 65°C, 15 min.
Precautions
1. For your safety and health, please wear lab coats and disposable gloves for operation.
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