Extended Data Figure 10. Measurement of ssDNA intermediates formed during BIR and analysis of BIR efficiency in the absence of Psf2 and Mcm4.

a. Schematic of the assay. b. Measurement of the relative increase of ssDNA at the indicated time after DSB induction compared to the amount of ssDNA in logarithmically growing cells (t=0). Measurement of ssDNA intermediate 10 and 40 kb from the site of strand invasion at the template chromosome and at a control locus on chromosome V which does not participate in recombination. c. An analysis of the growth of cells harboring temperature sensitive degron alleles of td-mcm4 and td-psf2. Both strains are inviable at 37°C even without overexpression of the ubiquitin ligase Ubr1. d. Western blot analysis of td-Mcm4 and td-Psf2 protein degradation. e. Southern blot analysis of the BIR assay in cells with conditional depletion of td-Mcm4 or td-Psf2. Quantification of the Southern blots is shown in Figure 2f.