5-Methyl-dCTP is a chemically modified deoxynucleotide analog in which the cytosine base contains a methyl group at the carbon-5 position Cytosine methylation at this position represents a frequent epigenetic modification observed extensively in mammalian genomic DNA During polymerase chain reaction (PCR) replacing conventional dCTP with 5-methyl-dCTP permits the synthesis of methylated DNA strands facilitating controlled in vitro studies of DNA methylation mechanisms Researchers employ this nucleotide analog to examine methylation-dependent recognition and cleavage activities of certain restriction and modification enzymes Additionally intracellular delivery of 5-methyl-dCTP can lead to its genomic incorporation altering epigenetic methylation patterns and associated gene expression profiles