Product Class: Other

dam Methyltransferase
NEBU recombinant 37 65 Heat SAM

M0222damMethyl_thumb

Product Introduction

  • Modifies the adenine residue (N6) of the sequence GATC
  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
Catalog # Size Concentration
M0222S 500.0 units 8000 units/ml
M0222L 2500.0 units 8000 units/ml

Product Information

Description

dam Methyltransferase modifies the adenine residue (N6) of the sequence GATC.

Product Source

An E. coli strain that carries plasmid pTP166 carrying the dam modification gene of E. coli
This product is related to the following categories:
Methyltransferases for Epigenetics,
Methyltransferases Products,
This product can be used in the following applications:
Methylated DNA Analysis 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 (dam-) Lambda DNA in 1 hour at 37°C in a total reaction volume of 10 µl against cleavage by MboI restriction endonuclease.

Reaction Conditions

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

1X dam Methyltransferase Reaction Buffer
50 mM Tris-HCl
5 mM β-ME
10 mM EDTA
(pH 7.5 @ 25°C)

Control Protection Assay

dam Methyltransferase is incubated with 1 µg of (dam-) Lambda DNA in 10 µl of 1X dam Methyltransferase Buffer, supplemented with 80 µM S-adenosylmethionine, for 1 hour at 37°C followed by 15 minutes at 65°C. The extent of protection is determined by addition of 40 µl 1X NEBuffer 3 supplemented with 10 mM MgCl2 and 10 units of MboI restriction endonuclease. Incubation at 37°C for 1 hour is followed by analysis on an agarose gel.

Storage Buffer

50 mM Tris-HCl
50 mM KCl
1 mM DTT
10 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

65°C for 20 minutes

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 dam Methyltransferase be heat inactivated?
  4. Does dam Methyltransferase require MgCl2?
  5. Is dam Methyltransferase sensitive to salt?
  6. Will DpnI cleave dam methylated DNA?
  7. Will any NEB methyltransferases (methylases) work on RNA?

Tech Tips

Always add the SAM to the reaction buffer just before doing the methylation reaction, because the SAM is unstable in an aqueous solution.

Keep the SAM frozen at -20C for a longer shelf life, fresh SAM is important for optimal methylation reactions.

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.

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