Product Class: Other

T3 RNA Polymerase
NEBU recombinant

Product Introduction

Bacteriophage T3 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T3 phage promoter. 

  • Isolated from a recombinant source
  • RNA probe preparation for hybridization
  • mRNA generation for in vitro translation systems
  • Supplied with 10X Reaction Buffer
  • Tested for the absence of other RNA polymerases, endonucleases, exonocleases and RNases
  • Getting ready to scale up RNA synthesis? Download our new technical note “Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production” for a generalized set of recommendations for synthesizing high yields of RNA.
Catalog # Size Concentration
M0378S 5000.0 units 50000 units/ml

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Product Information

Description

Bacteriophage T3 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T3 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T3 promoters. RNA produced using the T3 RNA Polymerase is suitable for many applications in research and biotechnology.

Product Source

Isolated from E. coli carrying a plasmid which contains T3 gene I.
This product is related to the following categories:
cDNA Synthesis & Reverse Transcriptases Products,
RNA Synthesis In vitro Transcription (IVT),
This product can be used in the following applications:
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0378S     -20    
      T3 RNA Polymerase M0378SVIAL -20 1 x 0.1 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 required to incorporate 1 nmol ATP into an acid-insoluble material in 1 hour at 37°C.

Reaction Condition
1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP and DNA template containing the T3 RNA Polymerase promoter. Incubate at 37°C.

Reaction Conditions

1X RNAPol Reaction Buffer

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

Storage Buffer

100 mM NaCl
50 mM Tris-HCl
1 mM EDTA
20 mM 2-mercaptoethanol
0.1% Triton® X-100
50% (w/v) Glycerol
pH 7.9 @ 25°C

Unit Assay Conditions

1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP and 2 μg T3 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 RNAprobes, the concentration of the radioactivenucleotide should be limited to 6 μM.

  2. To protect RNA against ribonuclease, RNaseinhibitor (NEB #M0314 or #M0307) should beadded to a final concentration of 1 U/μl.

  3. T3 RNA Polymerase is extremely sensitive to saltinhibition. The overall salt concentration shouldnot exceed 50 mM.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Standard RNA Synthesis (NEB #M0378)

Application Notes

FAQs & Troubleshooting

FAQs

  1. What is the promoter sequence of T3 RNA Polymerase?
  2. Does the transcription reaction with T3 RNA Polymerase require a primer?
  3. Does T3 RNA Polymerase leave an extra base at the end of a transcript?
  4. Will T3 RNA Polymerase work on uncut plasmid DNA?
  5. Can aberrant RNA be produced when using T3 RNA Polymerase?
  6. Can I use T3 RNA Polymerase to make high specific activity radiolabeled probes?
  7. What are the main causes of reaction failure using T3 RNA Polymerase?
  8. Why is the specific activity of the probe low?

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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