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. Author manuscript; available in PMC: 2021 Jul 20.
Published in final edited form as: Nature. 2021 Jan 20;590(7847):655–659. doi: 10.1038/s41586-020-03172-w

Extended Data Figure 3. BIR synthesis in pif1Δ, pol32Δ, and pol2-AID.

Extended Data Figure 3.

a, Strand invasion in pif1Δ and pol32Δ mutants 6h post DSB measured by DLC assay. DLC signal is normalized to ARG4 and compared to 0hr by fold changes. The means ± SD (n=3 independent biological repeats) are indicated. b, Kinetics of Ya disappearance in wt, pif1Δ, pol32Δ and pif1Δ pol32Δ. c, Schematic of primers used for AMBER analyses. d-f, AMBER analysis of BIR synthesis in wt (PIF1, POL32) (d), in pif1Δ (e), and pol32Δ (f). d-f each represents one out of three independent biological repeats that showed similar results (see Supplementary Table 6 for other repeats). Mean values of target to reference (ACT1) loci ratios were calculated by Poisson distribution based on 10,000 droplets with error bars representing upper and lower Poisson 95%CI. g, AMBER analysis of BIR synthesis following inactivation of Polε (pol2-degron) with schematic of analysis (top), degradation of AID-tagged Pol2 verified by Western Blot (middle) and calculation of copy number (bottom). The means ± SD (n=3 independent biological repeats) are indicated. * represents significant difference (p=0.0351) determined by t-test, two-tailed).