Product Class: Other

E. coli RNA Polymerase, Holoenzyme
Gray property icon for NEBU Gray property icon for incubation temperature 37°

Product Introduction

E. coli RNA Polymerase, Holoenzyme is the core enzyme saturated with sigma factor 70. The Holoenzyme initiates RNA synthesis from sigma 70 specific bacterial and phage promoters.

  • RNA synthesis from E. coli promoter
  • Transcription initiation studies
  • In vitro translation with PURExpress®
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Catalog # Size Concentration
M0551S 50 units 1000 units/ml

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Product Information

Description

E. coli RNA Polymerase, Holoenzyme is the core enzyme saturated with sigma factor 70. The Holoenzyme initiates RNA synthesis from sigma 70 specific bacterial and phage promoters.

E. coli RNA Polymerase, Core Enzyme consists of 5 subunits designated α, α, β´, β, and ω. The enzyme is free of sigma factor and does not recognize any specific bacterial or phage DNA promoters. The enzyme retains the ability to transcribe RNA from nonspecific initiation sequences. Addition of sigma factors will allow the enzyme to initiate RNA synthesis from specific bacterial and phage promoters. The core enzyme has a molecular weight of approximately 400 kDa.

Product Source

E. coli RNA Polymerase, Holoenzyme is isolated from E. coli strain BL21. The sigma factor 70 is purified from an E. coli strain carrying the cloned gene for sigma factor 70.
This product is related to the following categories:
RNA Modification,
Nucleotide Solutions for RNA,
RNA Synthesis In vitro Transcription (IVT),

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    M0551S -20
    Component Name Component # Stored at (°C) Amount Concentration
    E. coli RNA Polymerase, Holoenzyme M0551SVIAL -20 1 x 0.05 ml 1,000 units/ml
    E.coli RNA Polymerase Reaction Buffer B0550AVIAL -20 1 x 0.5 ml 5 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 1 nmol NTP into RNA in 10 minutes at 37°C.

Reaction Conditions

1X E.coli RNA Polymerase Reaction Buffer
Incubate at 37°C

1X E.coli RNA Polymerase Reaction Buffer
40 mM Tris-HCl
150 mM KCl
10 mM MgCl2
1 mM DTT
0.01% Triton® X-100
(pH 7.5 @ 25°C)

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.5 @ 25°C

Unit Assay Conditions

1X E. coli RNA Polymerase Reaction Buffer, supplemented with 0.5 mM of each NTP and 1 μg T7 phage DNA in 50 μl.

Application Features

  • RNA synthesis from E. coli promoter 
  • Transcription initiation studies
  • In vitro translation with PURExpress

FAQs & Troubleshooting

FAQs

  1. What is the difference between the E.coli RNA Polymerase, Core Enzyme and Holoenzyme?
  2. What are the major functions of these subunits?
  3. Can the Core and Holoenzyme be used in PURExpress?
  4. What are the other sigma factors in E.coli?

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.

Trademarks

New England Biolabs® and PUREXPRESS® are registered trademarks of New England Biolabs, Inc.
TRITON X-100™ is a trademark of The Dow Chemical Company.

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