Product Class: Other

Hhal Methyltransferase
neb31 cloned at NEB recombinant 37 No SAM

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

M0217HhaI_thumb

Product Introduction

HhaI Methyltransferase modifies the internal cytosine residue (C5) of the sequence GCGC

Catalog # Size Concentration
M0217S 1000.0 units 25000 units/ml

Product Information

Description

M.HhaI crystal (Xiaodong Cheng, Cold Spring Harbor Laboratory)
M.HhaI crystal (Xiaodong Cheng, Cold Spring Harbor Laboratory)

Product Source

An E. coli strain that carries the cloned HhaI modification gene from Haemophilus haemolyticus (ATCC 10014).
This product is related to the following categories:
Methyltransferases for Epigenetics,
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 HhaI 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

HhaI Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X rCutSmart 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 HhaI Methyltransferase is determined by the addition of 40 µl rCutSmart Buffer supplemented with 100 µg/ml BSA, 10 mM MgCl2 and 10 units of HhaI restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel.

Storage Buffer

50 mM Tris-HCl
150 mM NaCl
5 mM TCEP
0.1 mM EDTA
200 µg/ml BSA
pH 7.5 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 36994 daltons

Product Notes

  1. Storage of SAM: S-adenosylmethionine is stored at –20°C as 32 mM solution dissolved in sulfuric acid (0.005 M) and 10% ethanol. SAM in this solution stored under ideal conditions remains active for up to 6 months. SAM is unstable at (pH 7.5), 37°C, and should be replenished for reactions incubated longer than 4 hours. Many problems in achieving complete digestion can be alleviated by using fresh SAM.

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. What is the molecular weight of HhaI Methyltransferase?
  4. Will any NEB methyltransferases (methylases) work on RNA?
  5. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

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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