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Hhal Methyltransferase

Catalog # Concentration Size List Price Quantity Your Price
M0217S 25000 units/ml 1000 units $123.00
$110.70
Catalog # Size List Price Your Price
M0217S 1000 units $123.00
$110.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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

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).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Hhal Methyltransferase M0217SVIAL -20 1 x 0.04 ml 25,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
  S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM

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


Materials Sold Separately
Notes
  • 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
  • Hoffman, J.L. (1986). Biochemistry. 25, 4444-4449.
Quality Control Assay
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 Data Sheets
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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.

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