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SMCs might alter the topology of interacting DNA as a result of two processes: stoichiometric binding and dynamic supercoiling. Stoichiometric effects refer to DNA deformations constrained at the DNA interaction sites. Dynamic effects are cumulative alterations of DNA topology generated by a cyclic, energy-dependent activity. This chapter describes basic in vitro ensemble assays to measure the stoichiometric alterations of DNA topology caused by SMC binding and to test the potential DNA supercoiling generated by SMC activity. Lastly, it outlines gel electrophoresis conditions to reveal these changes.

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Gel Electrophoresis Analysis of SMC-Mediated Effects on DNA Topology

  • 实验方案

  • 发表日期：

    2026年

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  • Joaquim Roca

摘要

SMCs might alter the topology of interacting DNA as a result of two processes: stoichiometric binding and dynamic supercoiling. Stoichiometric effects refer to DNA deformations constrained at the DNA interaction sites. Dynamic effects are cumulative alterations of DNA topology generated by a cyclic, energy-dependent activity. This chapter describes basic in vitro ensemble assays to measure the stoichiometric alterations of DNA topology caused by SMC binding and to test the potential DNA supercoiling generated by SMC activity. Lastly, it outlines gel electrophoresis conditions to reveal these changes.