Product Class: Other

T7 RNA Polymerase
NEBU recombinant 37

GMP-grade reagent also available. Learn more.

Product Introduction

T7 RNA Polymerase is used for in vitro mRNA synthesis and is highly specific for the T7 phage promoter. Applications include:

Catalog # Size Concentration
M0251S 5000.0 units 50000 units/ml
M0251L 25000.0 units 50000 units/ml

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Product Information

Description

Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology. 

Reaction Conditions
1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP, and DNA template containing the T7 RNA Polymerase promoter. Incubate at 37°C. 

Product Source

Isolated from E.coli carrying a plasmid which contains the T7 RNA Polymerase gene. 
This product is related to the following categories:
RNA Synthesis In vitro Transcription (IVT)
This product can be used in the following applications:
Transcription-Mediated and NASBA Amplification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0251S     -20    
      T7 RNA Polymerase M0251SVIAL -20 1 x 0.1 ml 50,000 units/ml
      RNAPol Reaction Buffer B9012SVIAL -20 1 x 1 ml 10 X
  • M0251L     -20    
      T7 RNA Polymerase M0251LVIAL -20 1 x 0.5 ml 50,000 units/ml
      RNAPol Reaction Buffer B9012SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 μl in 1 hour at 37°C.

Reaction Conditions

1X RNAPol Reaction Buffer
Incubate at 37°C

1X RNAPol Reaction Buffer
40 mM Tris-HCl
6 mM MgCl2
1 mM DTT
2 mM spermidine
(pH 7.9 @ 25°C)

Storage Buffer

50 mM Tris-HCl
100 mM NaCl
20 mM β-ME
1 mM EDTA
50% Glycerol
0.1% (w/v) Triton® X-100
pH 7.9 @ 25°C

Unit Assay Conditions

1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP, and 1 µg T7 DNA in 50 µl.

Application Features

  • Radiolabeled RNA probes
  • Non-isotopic RNA labeling
  • Preparation of RNA vaccines
  • Guide RNA for gene targeting
  • mRNA for in vitro translation and micro injection 
  • RNA structure, processing and catalysis studies
  • RNA amplification
  • Anti-sense RNA for gene expression experiment 

Product Notes

  1. For radio labeled high specific activity RNA probes, the concentration of the radioactive nucleotide should be limited to 6 μM. 
  2. To protect RNA against ribonuclease, RNase inhibitor (NEB #M0314 or #M0307) should be added to a final concentration of 1 U/ μl. 
  3. T7 RNA Polymerase is extremely sensitive to salt inhibition. The overall salt concentration should not exceed 50 mM. 

References

  1. Schenborn, E.T. and Meirendorf, R.C. (1985). Nucl. Acids Res.. 13, 6223-6236.
  2. Davanloo, P., Rosenberg, A.H., Dunn, J.J. and Studier, F.W. (1984). Proc. Natl. Acad. Sci. USA. 81, 2035-2039.
  3. Sambrook, J., Fritsch, E.F. and Maniatis, T. Molecular Cloning: A Laboratory Manual, (2nd Ed.). 10.27-10.37.
  4. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd Ed.). 18.82-18.84.
  5. Melton, D.A., Kreig, P.A., Rebagliati, M.R., Maniatis, T., Zinn, K. and Green, M.R. (1984). Nucl. Acids Res.. 12, 7035-7056.
  6. Milligan, J.F., Groebe, D.R., Witherell, G.W. and Uhlenbeck, O.C. (1987). Nucl. Acids Res.. 15, 8783.
  7. Noren, C.J. et al. (1990). Nucl. Acids Res.. 18, 83-88.
  8. Kreig, P.A. and Melton, D.A. (1984). Nucl. Acids Res.. 12, 7057-7070.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Standard RNA Synthesis

Application Notes

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the promoter sequence of T7 RNA Polymerase?
  2. Is it possible to start transcription with an A?
  3. Does the transcription reaction with T7 RNA Polymerase require a primer?
  4. Does T7 RNA Polymerase leave an extra base at the end of a transcript?
  5. Will T7 RNA Polymerase work on single stranded substrate?
  6. Will T7 RNA Polymerase work on uncut plasmid DNA?
  7. Can aberrant RNA be produced when using T7 RNA Polymerase?
  8. Can I use T7 RNA Polymerase to make high specific activity radiolabeled probes?
  9. What are the main causes of reaction failure using T7 RNA Polymerase?
  10. Why is the specific activity of the probe low?
  11. How can the yield of RNA be maximized when using T7 RNA Polymerase?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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