library_construction_protocol: TrAEL-seq Extract Protocol:_x000B_Meiotic cultures (20 ml per strain) were harvested 5 hours into meiosis, pelleted, and washed twice with CHEF TE buffer (10 mM Tris-HCl, pH 7.5, 50 mM EDTA, pH 8.0). The cell pellet was resuspended in 300 µl CHEF TE, treated with 4 µl zymolyase (10 mg/ml), briefly vortexed, and incubated at 42°C for ~30 seconds. Low-melting-point agarose (1% SeaPlaque GTG in 125 mM EDTA, pH 8.0) was added, and the mixture was transferred into plug molds. After solidification on ice, plugs were transferred to LET buffer (10 mM Tris-HCl, pH 7.5, 0.5 M EDTA, pH 8.0) and incubated overnight at 37°C. Proteinase K treatment was performed in NDS buffer (10 mM Tris-HCl, pH 7.5, 0.5 M EDTA, 1% N-lauroylsarcosine) at 50°C overnight, followed by sequential washes with CHEF TE (1 hour), CHEF TE containing PMSF (two washes, 1 hour each), RNase T1 in CHEF TE (37°C for 1 hour), and a final wash with CHEF TE (1 hour). Plugs were stored at 4°C in CHEF TE until library preparation. TrAEL-seq Library Protocol:_x000B_Library preparation and sequencing were conducted as described in Kara et al., 2021.