Figure 5 .

Fluorescent crossover/noncrossover assay. (A) The configuration of P^GPD1-gfp* heteroalleles, NdeI restriction site polymorphisms, and flanking P^YKL050c-RFP and P^YKL050c-CFP markers is diagrammed. The relative frequency and position of DSBs are indicated with arrows. (B) DSBs at the ARG4::gfp* insertion site. A Southern blot of BglII-digested genomic DNA from a meiotic culture of a strain homozygous for sae2Δ and containing the fluorescence recombination reporter diagrammed in A. (C) FACS analysis of Gfp*⁺ recombinant tetrads. Approximately 1.6 × 10⁵ tetrads were analyzed. (D and E) Schematic and micrographs illustrating the configuration of markers when there is a noncrossover gene conversion (D) or either a gene conversion with an associated crossover or a crossover between the two gfp* mutations (E). (F) Tetrad with two Gfp*⁺ spores, possibly generated from premeiotic recombination or complex meiotic events. (G) MI nondisjunction, or (expected to be very rare) one class of four-strand double crossover. (H) Map distances (±SE) were calculated for the RFP–CFP interval in strains carrying the construct diagrammed in A. Statistical significance was evaluated by G test for the distribution of tetrad classes for each strain; spo11-HA and spo11-yf were compared to SPO11, whereas the SPO11 strain here was compared to SPO11 in Figure 3.