ue was divided by the 0-h value.
(2.8 MB PDF)
Figure S3. Quantile-Quantile Plots of IP:WCE Hybridization Ratios (log2) for Chromosome I from the Haploid Experiments (Strain GP6013) against Simulated Normal Values (with a Mean of 0 and Variance of 1)
Uninduced (0 h) data are from haploid experiment 2; induced (5 h) data are the mean of two haploid experiments. Normally distributed IP:WCE hybridization ratios (log2) should result in a straight line passing through the origin.
(A) 0 h, raw data.
(B) 0 h, filtered to remove isolated (single and double) linear values >2 (i.e., log2 > 1), identified as in Figure S2C. Note that these filtered data are nearly normally distributed.
(C) 5 h, raw data.
(D) 5 h, data filtered using criteria from Figure S2C. Note that the 5-h data have many values clearly above the normal distribution that persist after filtering, as expected for genuine Rec12-enrichment of DNA at linkage sites. The percent of total data with an abscissa value ≥1 is indicated for each plot.
(992 KB PDF)
Figure S4. Quantile-Quantile Plots of IP:WCE Hybridization Ratios (log2) for Chromosome I from the Diploid Experiment (Strain GP6203) against Simulated Normal Values (with a Mean of 0 and Variance of 1)
Normally distributed IP:WCE hybridization ratios (log2) should result in a straight line passing through the origin.
(A) 0 h, raw data.
(B) 0 h, filtered to remove isolated (single and double) linear values >2 (i.e., log2 > 1), identified as in Figure S2C. Note that these filtered data have some values above the normal distribution.
(C) 5 h, raw data.
(D) 5 h, data filtered using criteria from Figure S2C. Note that the 5-h data have many values clearly above the normal distribution that persist after filtering, as expected for genuine Rec12-enrichment of DNA at linkage sites. The percent of total data with an abscissa value ≥1 is indicated for each plot.
(1.8 MB PDF)
Figure S5. Distribution of Rec12-DNA Linkages in Two Haploid Inductions (Strain GP6013) and One Diploid Induction (Strain GP6203) Are Similar
Data for NotI fragment J are filtered and presented as in Figure S2C.
(1.9 MB PDF)
Figure S6. Southern Blot Analyses of DSBs in the 1.8-Mb Region at the Left End of Chromosome I
(A) Physical map and Southern blot hybridizations of NotI fragments encompassing 0.2–2 Mb from the left end of Chromosome I probed from the right (R) or left (L), as indicated. Haploid strain GP5411 (rad50S) was induced, and at the times indicated DNA was extracted and analyzed for DSBs by Southern blot hybridization as described by Young et al. .
(B) Coarse mapping of DSB positions from Figure S6A NotI blots.
(C) Finer scale mapping of the DSB sites identified in Figures S6A and S2C was carried out using DNA from haploid strain GP5411 and diploid strain GP3087 (rad50S), prepared as in Figure S6A. Also shown are blots bridging NotI fragments where breaks were observed in the bridging blots. One DSB site was refractory to analysis. The restriction enzyme used, its length, and the fragment end probed (R or L) are indicated. The DSBs are numbered as in Figures 3 and 4.
(1.3 MB PDF)
Figure S7. Rec12-DNA Linkages across the Entire S. pombe Genome
Data, filtered as in Figure S2C, are the IP:WCE ratios before induction (0 h; blue line) and 5 h after induction (red line; mean of two haploid experiments). Bars indicate the extent of the centromeric repeats. Carets indicate prominent (red) and weak (black) Rec12 peaks. Note that ∼0.5 Mb of rDNA, not shown, is at each end of Chromosome III.
(9.6 MB PDF)
Figure S8. Among Prominent Rec12 Peaks, Stronger Peaks Occur More Frequently in Large IGRs
Each prominent Rec12 peak was allocated to one of four bins based on its peak integral (i.e., strength). The percentage of all peaks in each bin that lie in IGRs >3 kb is indicated. Bin boundaries were chosen to allocate approximately equal numbers of peaks to each bin.
(11 KB PDF)
Figure S9. Meiotic DSB Positions and Relative Intensities are Similar in rad50+ and rad50S Backgrounds and in pat1+ and pat1–114 Backgrounds
S. pombe pat1–114 strains were meiotically in