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.
HhaI Methyltransferase modifies the internal cytosine residue (C5) of the sequence GCGC
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
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.
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Specifications & Change Notifications
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]
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]
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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.
