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. Author manuscript; available in PMC: 2014 Nov 14.
Published in final edited form as: EcoSal Plus. 2012 Nov;5(1):10.1128/ecosalplus.7.2.5. doi: 10.1128/ecosalplus.7.2.5

FIGURE 5.

FIGURE 5

In vitro assay for mismatch correction. The substrate supercoiled molecule contains a single GATC sequence which can be methylated on either the viral (V) or complementary (C) strand (+/− or −/+) or both (+/+) or neither (−/−). A control molecule (0/0) contains a GATT sequence in place of GATC. The substrate also contains a T-G mismatch (carats) in the indicated sequence which is part of both a HindIII and an XhoI recognition site. Upon completion of the in vitro reaction in extracts or with purified components, the substrate DNA is digested with ClaI, HindIII and XhoI. Correction of the mismatch will lead to resistance to one of the restriction enzymes. The substrate without the GATC (0/0) or a fully methylated GATC (+/+) shows no repair products while the hemi-methylated (+/− or −/+) or unmethylated substrates (−/−) show repair products derived preferentially from one strand. The Figure has been modified from reference (88).