Data from: Polymicrobial Lysis-Hi-C experiments of three bacteria and plasmids derived from HiC-Pro

<p>Microbes interact in biofilms and polymicrobial infections in a spatially structured manner, yet next-generation sequencing approaches generally fail to recover <em>in situ</em> spatial proximity between distinct genotypes in the interactome or eDNA matrix. This study explored a modified Hi-C approach involving an initial lysis phase prior to DNA cross-linking, to test whether adjacent cell chromatin can be cross-linked, anticipating that this could provide a new avenue for the study of spatial-mutational dynamics in structured microbial communities. An artificial polymicrobial mixture of <em>Pseudomonas aeruginosa</em>, <em>Staphylococcus aureus</em>, and <em>Escherichia coli</em> was lysed for 1 to 18 hours, then prepared for Hi-C. A murine biofilm infection model was treated with sonication, mechanical lysis, or chemical lysis before Hi-C. Bioinformatic analyses of resulting Hi-C interspecies chromatin links showed that while microbial species differed from one another, generally lysis significantly increased links between species and increased the distance of Hi-C links within species, while also increasing novel plasmid-chromosome links. The success of this modified lysis-Hi-C protocol in creating extracellular DNA links is a promising first step toward a new lysis-Hi-C-based method to recover genotypic microgeography in polymicrobial communities, with potential future applications in diseases with localized resistance, such as cystic fibrosis lung infections and chronic diabetic ulcers.</p>