Product Class: Other

Hi-T7® RNA Polymerase (High Concentration)
NEBU 50

Product Introduction

Hi-T7 RNA Polymerase is designed for in vitro transcription of RNA at higher temperatures.

  • Active from 37-56°C, optimal incubation temperature is 50-52°C.
  • In vitro transcription using T7 phage promoter
  • High concentration version of Hi-T7 RNA Polymerase (NEB #M0658)
  • Increased co-transcriptional capping efficiency with cap analogs
  • Decreased immunogenicity from RNA synthesized at higher temperature due to reduced dsRNA by-product formation
  • Recommended for experienced users with knowledge of transcription optimization 
  • Hi-T7 RNA Polymerase Reaction buffer and magnesium chloride included
  • 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
M0470T 50000 units 1000000 units/ml

Product Information

Description

Hi-T7 RNA Polymerase (High Concentration) is offered at a 20-fold higher concentration than our standard Hi-T7 RNA Polymerase (NEB #M0658) and is ideal for experienced users interested in building and optimizing their own in vitro transcription reactions. The enzyme is accompanied by Hi-T7 RNA Polymerase Reaction Buffer and Magnesium Chloride Solution to further enable testing of different conditions such as nucleotide concentrations. 

In vitro transcription involves multiple components in addition to the RNA Polymerase including ribonucleotides, inorganic pyrophosphatase and RNase Inhibitor, which can be purchased separately. Please refer to the Related Products section for details. 

Hi-T7 RNA Polymerase is an engineered DNA-dependent RNA polymerase that is highly specific for T7 phage promoters and is designed to function at higher temperatures than the wild-type bacteriophage T7 RNA Polymerase. Hi-T7 RNA Polymerase functions at an optimal temperature of 50-52°C.

Product Source

An E.coli strain that carries an engineered His-tag 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
  • M0470T     -20    
      Hi-T7® RNA Polymerase (High Concentration) M0470TVIAL -20 1 x 0.05 ml 1,000,000 units/ml
      Hi-T7® RNA Polymerase Reaction Buffer B0658AVIAL -20 1 x 1 ml Not Applicable
      MgCl2 Solution B2534AVIAL -20 1 x 0.5 ml 0.2 M

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 1 nmol of ATP into acid-insoluble material in 1 hour at 50°C

Reaction Conditions

1X Hi-T7® RNA Polymerase Reaction Buffer
Incubate at 50°C

1X Hi-T7® RNA Polymerase Reaction Buffer
40 mM Tris-HCl
4 mM MgCl2
50 mM NaCl
2 mM spermidine
1 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

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

Unit Assay Conditions

1X Hi-T7 RNA Polymerase Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP and 1 µg T7 phage DNA in 50 µl reaction volume.

Application Features

  • mRNA for in vitro translation, microinjection and transfection
  • Radiolabeled RNA probes
  • Non-isotopic RNA labeling
  • Guide RNA for gene targeting
  • RNA structure, processing and catalysis studies
  • Anti-sense RNA for gene expression
  • RNA amplification
  • Isothermal amplification

Protocols, Manuals & Usage

Protocols

  1. Protocol for Standard RNA Synthesis
  2. Purification of IVT RNA

Application Notes

FAQs & Troubleshooting

FAQs

  1. Can I use a different reaction buffer for in vitro transcription with Hi-T7 RNA Polymerase?
  2. Does Hi-T7 RNA Polymerase recognize the same promoter sequence as the wild-type T7 RNA Polymerase from bacteriophage?
  3. At what temperature is Hi-T7 RNA Polymerase active?
  4. What are the main causes of reaction failure when using Hi-T7 RNA Polymerase?
  5. What templates can be used for in vitro transcription with Hi-T7 RNA Polymerase?
  6. Can Hi-T7 RNA Polymerase transcribe off of an uncut plasmid?

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.