Product Class: Other

HaeIII Methyltransferase
NEBU cloned at NEB recombinant 37 65 Heat SAM

The L size of this product has been discontinued on 11/15/16. The S size will still be available.

M0224HaeIIImethyl_thumb

Product Introduction

HaeIII Methyltransferase modifies the internal cytosine residue (C5) of the sequence GGCC.

This enzyme comes from an E. coli strain that carries the cloned HaeIII modification gene from Haemophilus aegyptius (ATCC 11116).

Catalog # Size Concentration
M0224S 500.0 units 10000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the cloned HaeIII modification gene from Haemophilus aegyptius (ATCC 11116).
This product is related to the following categories:
Methyltransferases for Epigenetics Products,
Methyltransferases Products,

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 HaeIII restriction endonuclease.

Reaction Conditions

1X HaeIII Methyltransferase Reaction Buffer
Supplement with 80 µM S-adenosylmethionine (SAM)
Incubate at 37°C

1X HaeIII Methyltransferase Reaction Buffer
50 mM Tris-HCl
50 mM NaCl
10 mM DTT
(pH 8.5 @ 25°C)

Control Protection Assay

HaeIII Methyltransferase is incubated with 1 µg of λ DNA in 10 µl of 1X HaeIII Methyltransferase Reaction Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection is determined by the addition of 40 µl NEBuffer 2.1 and 10 units of HaeIII restriction endonuclease. Incubation for 1 hour at 37°C is followed by analysis on an agarose gel.

Storage Buffer

50 mM Tris-HCl
1 mM DTT
10 mM EDTA
200 µg/ml BSA
50% Glycerol
50 mM KCl
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Product Notes

  1. HaeIII Methyltransferase protects DNA against cleavage by NotI.

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

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. Does HaeIII Methyltransferase require DTT?
  4. Can HaeIII Methyltransferase be heat inactivated?
  5. Does HaeIII Methyltransferase require MgCl2?
  6. What is the molecular weight of HaeIII Methyltransferase?
  7. 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.

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.