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Smc5/6 is an essential genome maintenance complex that regulates DNA replication and repair. Its roles in managing DNA intermediates generated from these processes are intimately linked to its ability to associate with various forms of DNA. To gain insights into how Smc5/6 engages with different DNA structures, we use the C-Trap single-molecule setup in combination with several DNA substrates. We will presence our recent findings of how Smc5/6 preferentially associates with DNA junctions flanking a ssDNA gap, a common DNA intermediate generated during genome maintenance processes. We will also discuss how other genome protecting factors can influence Smc5/6 junction binding behaviors and the implications of our findings in genome maintenance processes.
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.