Product Class: Other

EcoRI Methyltransferase
neb31 cloned at NEB recombinant 37 65 Heat SAM

EcoRI Methyltransferase is now supplied with rCutSmart Buffer (12/1/2021) 

M0211EcoRIMethyl_thumb

Product Introduction

  • EcoRI Methyltransferase modifies the internal adenine residue (N6) of the sequence GAATTC
  • Supplied with rCutSmart Buffer and 32 mM S-adenosylmethionine (SAM)
Catalog # Size Concentration
M0211S 10000.0 units 40000 units/ml

Product Information

Description

EcoRI Methyltransferase modifies the internal adenine residue (N6) of the sequence GAATTC.

Product Source

An E. coli. strain that carries the cloned EcoRI modification gene from Escherichia coli RY13 (R. N. Yoshimori).
This product is related to the following categories:
Methyltransferases for Epigenetics Products,
Methyltransferases Products,
This product can be used in the following applications:
Restriction Enzymes for Epigenetics

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to protect 1 μg λ DNA in 1 hour at 37°C in a total reaction volume of 10 μl against cleavage by EcoRI restriction endonuclease

Reaction Conditions

1X rCutSmart™ Buffer
Supplement with 80 µM S-adenosylmethionine (SAM)
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Control Protection Assay

rCutSmart Buffer is incubated with 1 µg of λ DNA in 10 µl 1X EcoRI Methyltransferase Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection by EcoRI Methyltransferase is determined by the addition of 40 µl rCutSmart Buffer supplemented with 10 mM MgCl2 and 5 units of EcoRI restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel.

Storage Buffer

100 mM KPO4
200 mM NaCl
10 mM β-ME
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Molecular Weight

Theoretical: 37911 daltons

Product Notes

  1. EcoRI Methyltransferase is inhibited by MgCl2. Only 50% activity is retained at a concentration of 4 mM MgCl2.

References

  1. Hoffman, J.L. (1986). biochemistry. 25, 4444-4449.

Protocols, Manuals & Usage

Protocols

  1. Recommended Protocol for 3H labeling of DNA
  2. Recommended Protocol for Methylation of Genomic DNA

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. Is S-adenosylmethionine (SAM) supplied with the Methyltransferase?
  2. What should be considered if the methylation is not going to completion?
  3. Can EcoRI Methyltransferase be heat inactivated?
  4. Does EcoRI Methyltransferase require MgCl2?
  5. Will EcoRI methylation block the ApoI restriction enzyme?
  6. Will any NEB methyltransferases (methylases) work on RNA?

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.

Specification Change Notifications

EcoRI Methyltransferase is now supplied with rCutSmart Buffer (12/1/2021)

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 [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.

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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