FAQ: Are the Lambda DNA (NEB #N3011) and Lambda DNA (dam-) (NEB #N3013) methylated?

Lambda DNA N3011 is methylated but we do not know exactly which A and C sites get methylated, since the methylation in Lambda is not 100%. The DNA is extracted from a dam+, dcm+, EcoK+ E.coli (strain C190).


NEB also offers Lambda DNA (dam-) (
NEB #N3013), which is grown in a E.coli strain which is dam-, dcm+, EcoK+ (strain GM33).

The methylase encoded by the dam gene (Dam methylase) transfers a methyl group to the N6 position of the adenine residues in the sequence GATC (1,2). The Dcm methylase, encoded by the dcm gene, methylates the internal cytosine residues in the sequences CCAGG and CCTGG (1,3) at the C5 position. The EcoKI methylase, M.EcoKI, modifies adenine residues in the sequences AAC(N6)GTGC and GCAC(N6)GTT. For more information please see:
https://www.neb.com/tools-and-resources/selection-charts/dam-dcm-and-cpg-methylation

Not all DNA isolated from E.coli is methylated to the same extent. While pBR322 DNA is fully modified, only about 50% of Lambda DNA dam sites are methylated, presumably because the methylase does not have the opportunity to methylate the DNA fully before it is packaged into the phage head.

1. Marinus, M.G. and Morris, N.R. (1973) J. Bacteriol. 114, 1143–1150. PMID: 4576399
2. Geier, G.E.and Modrich, P. (1979) J. Biol. Chem. 254, 1408–1413. PMID: 368070
3. May, M.S. and Hattman, S.(1975) J. Bacteriol. 123, 768–770. PMID: 1097428