Product Class: Other

T7 RNA Polymerase
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Item# Description List Price Web Price Qty
M0251L T7 RNA Polymerase - 25,000 units $400.00
$360.00
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M0251S T7 RNA Polymerase - 5,000 units $100.00
$90.00
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Product Introduction

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

  • Radiolabeled RNA probe preparation
  • RNA generation for studies of RNA structure, processing and catalysis
  • Expression control via anti-sense RNA
  • mRNA, sgRNA synthesis
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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. 

Advantages and Features

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 

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 Conditions

    50 mM Tris-HCl
    100 mM NaCl
    20 mM β-ME
    1 mM EDTA
    50% Glycerol
    0.1% 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.

    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

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)

    Tools & Resources

    Selection Charts

    • DNA Polymerase Selection Chart
    • RNA Polymerase Selection Chart

    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. How can the yield of RNA be maximized when using T7 RNA Polymerase?
    9. Can I use T7 RNA Polymerase to make high specific activity radiolabeled probes?
    10. What are the main causes of reaction failure using T7 RNA Polymerase?
    11. Why is the specific activity of the probe low?

    Troubleshooting

    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

    This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

    While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

    For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

    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.