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Evaluation and quality control of LSHase activity. Substrates of 3.1 kb that mimic reannealed hybrid restriction fragments, either perfectly complementary (IBD) or with a single base pair mismatch (N-Id), and either as covalently closed circular (ccc) or linear (lin) forms, were incubated with LSHase and/or exo III (exo) and treated with the DAP procedure (resin), as indicated. Agarose gels of reaction products were stained with ethidium bromide. Lane M – high mass ladder, Invitrogen.
