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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.