Epigenetics

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【Zymo】

Epigenetics>【Zymo】

	CPG甲基转移酶（E2010）

CpG Methylase (M.SssI)---CPG甲基转移酶

About CpG Methylase (M.SssI)

The CpG Methylase from Zymo Research completely methylates all cytosines (C5) in double-stranded, non-methylated and hemimethylated DNA having the dinucleotide sequence 5'…CpG…3'. The recombinant methylase is isolated from an E. Coli strain that expresses the methyltransferase gene from Spiroplasma sp. strain MQ1. Reaction conditions have been optimized to maximize the processivity of the enzyme to ensure rapid, complete, and reproducible methylation of DNA for accurate DNA methylation analysis. This product is supplied with 10X Reaction Buffer and S-adenosylmethionine cofactor.

Format	Provided in solution (4 units/µl) w/ 10X Reaction Buffer and 20X SAM
Source	Recombinant methylase is isolated from E. Coli expressing methyltransferase gene from Spiroplasma sp. strain MQ1.
Storage	Store at -20°C for up to 12 months. Avoid repeated freeze/thawing of reagents. Prolonged storage is at ≤ -70°C.
Enzyme Commission Number	(EC 2.1.1.37)
Unit Definition	One unit is defined as the amount of enzyme required to "protect" 1 µg of λ DNA against cleavage by BstUI restriction endonuclease in a total reaction volume of 20 µl for 1 hour at 37°C.
Reaction Conditions	CpG Methylase in 1X CpG Reaction Buffer w/ 600 µM SAM. Incubate reaction mixtures at 30°C (Performing the Methylase reaction at 30°C rather than 37°C promotes optimal reaction kinetics).
Inactivation	Heat-inactivate the enzyme at 65°C for 20 minutes.
Standard Reaction Setup	The reaction volumes can be adjusted accordingly depending on experimental requirements (see protocol).

The CpG Methylase from Zymo Research catalyzes complete methylation of the CpG sites in DNA. Methylase activities of CpG Methylase from Zymo Research versus that of another supplier were tested for complete methylation of equivalent amounts of a linearized plasmid DNA using reaction conditions recommended by the supplier. Completion of CpG methylation was assessed by resistance to digestion with a methylation-specific endonuclease (HpaII) and subsequently analyzed in an agarose gel. As shown in the figure above, the CpG Methylase from Zymo Research completely methylated the CpG sites in the DNA whereas that of the other supplier did not. Samples were assayed in duplicate.

For complete, in vitro methylation of DNA for methylation analysis.

Methylation of chromatin DNA for DNA accessibility studies.

Inhibition of endonucleases with overlapping CpG sequence recognition.

[3H]-labeling of DNA.