T3 RNA Polymerase

Product Information

• Reagents Supplied

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	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.

• 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

• Related Products

  Companion Products

  • RNA Loading Dye, (2X)
  • RNase Inhibitor, Human Placenta
  • RNase Inhibitor, Murine
  • Pyrophosphatase, Inorganic (E. coli)
  • Low Range ssRNA Ladder
  • ssRNA Ladder
  • Vaccinia Capping System
  • mRNA Cap 2 O Methyltransferase
  • E. coli Poly(A) Polymerase
  • Ribonucleotide Solution Mix
  • Ribonucleotide Solution Set
  • SP6 RNA Polymerase
  • T7 RNA Polymerase

• 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 https://www.neb.com/en/#M0314 or https://www.neb.com/en/#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 https://www.neb.com/en/#M0378)

• Application Notes

  • Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production

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?

Citations & Technical Literature

• Citations

  Product Citation Tool

Quality, Safety & Legal

• Quality Control Assays

  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 & Change Notifications

  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]

  • M0378S_v1

• 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]

  • M0378S_v0_10008846
  • M0378S_v1_10008846
  • M0378S_v1_10031654
  • M0378S_v1_10060197
  • M0378S_v1_10085499
  • M0378S_v1_10110336
  • M0378S_v1_10124268
  • M0378S_v1_10152089
  • M0378S_v1_10204341
  • M0378S_v1_0011712
  • M0378S_v1_10239122

• Safety Data Sheets

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

  • T3 RNA Polymerase
  • RNAPol Reaction Buffer

• 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.