2.1.1.72 | -999 |
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the enzyme catalyzes the methylation of adenine at the N-6 position within its DNA recognition sequence GATC. Enzymatic methylation of the hemimethylated GATC site results in destabilisation of the duplex, increasing the proportion of dissociated DNA, and producing an observable increase in fluorescence. By using a suitable oligonucleotide substrate, labeled on adjacent strands with a fluorophore (fluorescein) and quencher (dabcyl), the destabilization of duplex DNA due to adenine methylation can be monitored by fluorescence measurements. The separation of fluorophore and quencher during strand dissociation causes an observable increase in fluorescence, providing a reproducible, direct, and real-time activity assay |
744009 |