MscI is blocked by overlapping dcm methylation if the site TGGCCAgg or ccTGGCCA occurs. To avoid dcm methylation, use a dcm- deficient strain for cloning such as dam-/dcm- Competent E.coli (#C2925). For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE.