GpC Methyltransferase (M.CviPI)

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0227S     -20       GpC Methyltransferase (M.CviPI) M0227SVIAL -20 1 x 0.05 ml 4,000 units/ml   GC Reaction Buffer B0227SVIAL -20 1 x 1.5 ml 10 X   S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM
M0227L     -20       GpC Methyltransferase (M.CviPI) M0227LVIAL -20 1 x 0.25 ml 4,000 units/ml   GC Reaction Buffer B0227SVIAL -20 1 x 1.5 ml 10 X   S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM

Application Features

• Blocking restriction endonuclease cleavage
• Altering the physical properties of DNA
• Uniform [³H]-labeling of DNA

Materials Sold Separately

• S-adenosylmethionine (SAM)

1. • S-adenosylmethionine (SAM) is supplied as a 32 mM solution in 0.005 M sulfuric acid and 10% ethanol. Under these conditions SAM is stable for up to 6 months when stored at -20°C.
   • SAM is unstable at (pH 7.5), 37°C, and should be replenished in reactions incubated longer than 4 hours.
2. Requires fresh DTT for optimum activity. For best results, mix assay buffer fresh for each use.
3. Methylation at cytosine residues has also been shown to affect the physical properties of DNA, including lowering the free energy of Z-DNA formation (1), increasing the helical pitch of DNA (2), and altering the kinetics of cruciform extrusion (3). Positions of 5-methylcytosine can be identified due to decreased reactivity to hydrazine in chemical sequencing protocols (4).
4. MgCl₂ is not required as a cofactor.

1. Zacharias, W. et al. (1988). Biochemistry. 27, 2970-2978.
2. Gruenbaum, Y. et al. (1982). Nature. 295, 620-621.
3. Murchie, A.I. and Lilley, D.M. (1989). J. Mol. Biol.. 205, 593-602.
4. Ohmori, H. et al. (1978). Nucl. Acids. Res. 5, 1479-1485.
5. Xu, S. et al. (1998). Nucl. Acids Res.. 26, 3961-3966.
6. Kladde, M.P. et al. (1991). Methods Enzymol. . 304, 431-447.

1. Recommended Protocol for ³H labeling of DNA
2. Recommended Protocol for Methylation of Genomic DNA

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

1. Will GpC Methyltransferase (M. CviPI) work in rCutSmart buffer?
2. Will any NEB methyltransferases (methylases) work on RNA?

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.

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]

• M0227S_L_v2

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]

• M0227S_v1_10008188
• M0227L_v1_10008189
• M0227L_v1_10027571
• M0227L_v1_10030907
• M0227S_v1_10030073
• M0227L_v1_10032146
• M0227L_v1_10037014
• M0227S_v1_10038535
• M0227L_v1_10039368
• M0227L_v1_10042755
• M0227L_v1_10044283
• M0227S_v1_10047062
• M0227L_v1_10048558
• M0227L_v1_10051499
• M0227S_v1_10048744
• M0227L_v1_10055155
• M0227L_v1_10060523
• M0227S_v1_10064033
• M0227L_v1_10064030
• M0227L_v1_10068227
• M0227L_v2_10084262
• M0227L_v2_10086962
• M0227S_v2_10084260
• M0227L_v2_10094825
• M0227L_v2_10097608
• M0227L_v2_10102104
• M0227L_v2_10106595
• M0227L_v2_10106687
• M0227L_v2_10115560
• M0227L_v2_10116063
• M0227S_v2_10116064
• M0227L_v2_10126547
• M0227L_v2_10128991
• M0227L_v2_10132529
• M0227S_v2_10145043
• M0227L_v2_10148518
• M0227L_v2_10162752
• M0227L_v2_10164704
• M0227L_v2_10171156
• M0227S_v2_10167440
• M0227L_v2_10184697
• M0227L_v2_10186025
• M0227S_v2_10191833
• M0227L_v2_10192494
• M0227S_v2_10203679
• M0227L_v2_10208855
• M0227S_L_v1_0021709
• M0227S_L_v1_0041804
• M0227L_v2_10219527
• M0227L_v2_10224836
• M0227S_v2_10224835

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• GpC Methyltransferase (M.CviPI)
• GC Reaction Buffer
• S-adenosylmethionine (SAM)

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.

Licenses

Licensed under U.S. Patent 7,034,116 and 6,492,168.
Licensed under U.S. Patent 6,492,168 and 7,034,116