EcoGII Methyltransferase is a non-specific methyltransferase that modifies adenine residues (N6) in any sequence context.
- Supplied with 10x CutSmart® Buffer and 32 mM S-adenosylmethionine (SAM)
- This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.
EcoGII Methylation of Various Substrates
Substrate % Methylation* DNA Substrates1 Plasmid DNA >75% Genomic DNA >50% ds DNA oligo >50% ss DNA oligo >50% DNA/RNA hybrid >50% RNA Substrates2 Total RNA 5-10% mRNA (in vitro transcribed) 5-10% DNA/RNA hybrid 5-10%
*Methylation Conditions: Methylation of 1 µg substrate with 5 U EcoGII Methyltransferase in 1X CutSmart® Buffer supplemented with 160 µM SAM for 60 minutes at 37°C. Following methylation, samples were column purified, digested to nucleosides and analyzed by LCMS to determine the ratio of N6mA to A (% methylation).
1 Tested substrates include: plasmid DNA (pBR322 #N3033, pUC19 #N3041), genomic DNA (Human Embryonic Kidney (HEK293) cells, Mouse Ear and Tail, Rat Liver and Kidney, N6-methyladenine-free λ DNA #N3013), ds DNA oligo (80 nucleotides), ss DNA oligo (60 nucleotides), DNA/RNA hybrid (80 nucleotides), total RNA (HeLa cell RNA, Mouse Kidney RNA, Rat Brain RNA), mRNA (in vitro transcribed 1.8 kb FLuc mRNA).
2 Methylation levels of >20% have been observed by using less substrate (250 ng) and increasing the amount of enzyme (15 U).
Methylation levels can be increased by decreasing the amount of substrate, increasing the amount of enzyme and/or increasing the length of the methylation reaction.
Product SourceAn E. coli strain that carries the cloned EcoGII methyltransferase gene from E. coli (strain C227-11)(1).
The following reagents are supplied with this product:
Store at (°C) Concentration CutSmart® Buffer -20 10 X S-adenosylmethionine (SAM) -20 32 mM
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