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Cat. No.: MMPEI-50 (for 50U)

Cat. No.: MMPEI-200 (for 200U)

Cat. No.: MMPEI-1k (for 1000U)

Description

CpG Methyltransferase (M.MpeI), abbreviated as M.MpeI and also known as DNA methyltransferase from M. penetrans or M.MpeI CpG methyltransferase, is a CpG methyltransferase derived from Mycoplasma penetrans. It is expressed and purified from Escherichia coli. This enzyme shares the same enzymatic function as CpG Methyltransferase (M.SssI) (CpG Methyltransferase from Spiroplasma sp. strain MQ1), recognizing all cytosine residues (C5) within the 5'-CG-3' sequence and performing methylation modifications.

M.MpeI serves as an essential tool enzyme in the field of epigenetics. It is widely used to simulate methylation modifications found in eukaryotic genomes, aiding in the exploration of cytosine methylation functions and related research applications.

Activity Definition

One unit of the enzyme is defined as the amount required to protect 1 μg of λDNA from cleavage by BstUI restriction endonuclease within a total reaction volume of 20 μl at 37°C in 1 hour.

Storage

Store at -20ºC for at least two years.

Precautions

• CpG Methyltransferase (M.MpeI) (4U/µl) should be kept on ice during use and returned to -20ºC immediately after use.
• For M.MpeI (20U/µl), if high enzyme concentration is not required, it can be diluted to 4U/µl using the provided M.MpeI Storage Buffer.
• When conducting in vitro methylation, consider the high density of CpG dinucleotide sites in the DNA substrate. For instance, λDNA (48,502 bp) contains 3,112 CpG sites, meaning that in a solution of 0.1 mg DNA/ml, the methyltransferase methyl acceptor sites are at a concentration of 19 µM. This is significant as the recommended methyl donor concentration is 160 µM, eight times higher than the acceptor sites. Reducing DNA concentration to less than 0.02 mg/ml ensures an adequately high methyl donor concentration to drive the reaction while also minimizing the potential inhibition caused by S-adenosyl-L-homocysteine (AdoHcy), a byproduct of the reaction.
• This product is intended for use only by trained professionals for scientific research. It must not be used for clinical diagnosis or treatment, nor for food or drug applications. Storage in residential areas is not permitted.
• For your safety and health, always wear a lab coat and disposable gloves during handling.