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Structural Maintenance of Chromosome (SMC) complexes, cohesin, condensin and Smc5/6, play a fundamental role in genome organization, facilitating chromosome compaction, segregation, and DNA repair. Despite their essential functions, the mechanisms by which the complexes interact with different DNA substrates and influence topological transitions remain not fully understood. Using the LUMICKS C-Trap, we have employed single-molecule approaches to analyze the behavior of purified SMC complexes, focusing on yeast cohesin and human SMC5/6, on different DNA substrates, including double-stranded (dsDNA) and single-stranded DNA (ssDNA). Additionally, we have used a quadrupole optical trap to bridging by SMC complexes and their effect on DNA decatenation by Topoisomerase IIα (Top2A). These findings provide new insights into the fundamental properties and requirements of cohesin and Smc5/6’s interaction with DNA substrates, as well as their ability to bridge two independent DNAs.
Precisely manipulating genetic material at the single molecule level is gaining importance across life sciences – and so do the tools that allow researchers to do exactly that. The C-Trap system combines single molecule fluorescence microscopy with optical tweezers to manipulate DNA, allowing researchers to directly observe and track molecular events as they occur. Designing and creating specific DNA constructs is crucial for maximizing the potential of single molecule studies. In this application note we introduce the powerful combination of cutting edge biochemistry and single-molecule visualization methods to increase throughput and maximize the results gained from each individual measurement.