100 OD of cells were mixed a synthetic DNA:RNA hybrid spike-in, in a 1:1 ratio with cell count, and 5% of C. glabrata cells prior to lysis. Genomic DNA was phenol-extracted and isolated by ethanol precipitation. DNA:RNA hybrids were then immunoprecipitated using the S9.6 antibody. Library were created using the xGen ssDNA & Low-Input DNA Library Prep Kit (Integrated DNA Technologies) according to the manufacturer instructions, with 16 cycles of amplification. Post-amplification libraries were size selected at 350-1000bp using SPRIselect beads (Beckman Coulter). Libraries were further quantified by Qubit and Tapestation (Agilent). ssDNA-seq