T7 RNA Polymerase
| Cat. No. | E041 | ||||||
| Name | T7 RNA Polymerase | ||||||
| Unit | 100 ul (5000 U) | ||||||
| Category | Molecular Biology Enzymes and Kits | ||||||
| Description |
T7 RNA Polymerase is a DNA-dependent enzyme that catalyzes the synthesis of RNA in the 5' to 3' direction. It works exclusively in the presence of its T7 phage promoter sequence, ensuring high specificity for transcription. This enzyme does not recognize SP6 or T3 RNA Polymerase promoter sequences, making it ideal for applications requiring precise and controlled RNA production from T7-driven templates.
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| Application |
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| Concentration | 50 U/ul | ||||||
| Components | Enzyme supplied with 10X Reaction Buffer | ||||||
| Storage Buffer | 50 mM Tris-HCl (pH 8), 1 mM EDTA, 20 mM β-ME, 100 mM NaCl, 0.1% Triton®X-100 and 50% (v/v) Glycerol. | ||||||
| Storage Condition | Store all components at -20°C. | ||||||
| Note |
One unit is defined as the amount of T7 RNA Polymerase that is required to incorporate 1 nmol ATP into acid-insoluble material in a 50 μl reaction volume in 1 hour at 37°C in 1X T7 RNA Polymerase Reaction Buffer. |
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. E041 |
| What is T7 RNA Polymerase used for? | |
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T7 RNA Polymerase is used for synthesizing RNA transcripts, which can be used as hybridization probes, for in vitro translation, or for generating biologically active mRNA. It is also useful for creating large amounts of labelled or non-labelled RNA.
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| What is the optimal reaction condition for T7 RNA Polymerase? | |
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The recommended reaction condition is to use 1X T7 RNA Polymerase Reaction Buffer and incubate the reaction at 37°C. Ensure the reaction mixture includes fresh dithiothreitol (DTT) and maintain the salt concentration below 50 mM for optimal enzyme activity.
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| How do I increase the yield of RNA in my reaction? | |
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To increase RNA yield, you can increase the concentration of NTPs (up to 4 mM each) and adjust the magnesium concentration to 4 mM above the total NTP concentration.
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| What is the definition of one unit of T7 RNA Polymerase? | |
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One unit of T7 RNA Polymerase is defined as the amount required to incorporate 1 nmol ATP into acid-insoluble material in a 50 μl reaction volume over 1 hour at 37°C in 1X T7 RNA Polymerase Reaction Buffer.
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| Why is DTT important for T7 RNA Polymerase activity? | |
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Dithiothreitol (DTT) is essential for maintaining the enzyme’s activity. If you notice a decrease in activity, it may be due to the breakdown of DTT over time, even when stored at -20°C. In such cases, you should supplement the reaction with fresh DTT at a final concentration of 10 mM.
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| What is the source of this enzyme? | |
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The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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