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Table S1 PCR conditions for amplifying microsatellites in (A) Columbia spotted frogs and (B) long-toed salamanders. PCR profile was: initial denaturing 95 °C for 15 min, followed by 35–38 cycles of 95 °C for 30 s, annealing temperature for 90 s, and 72 °C for 60 s, followed by final extension of 60 °C for 30 min. Touchdowns were staged in increments of 0.5 °C. All reactions used 1X Qiagen Multiplex Master Mix. Q indicates 5X Qiagen Q solution

Fig. S1 Population ancestry values derived from STRUCTURE analyses (Prichard et al. 2000), indicating the most probable number of genetic groups using the ΔK statistic. Individual-based measurements yield very similar results. We conducted 10 simulations for each number of distinct genetic groups (K), ranging from 1 to 10, used the admixture model, and assumed correlated allele frequencies (Falush et al. 2003). The burn-in length was 100 000 and the run length was 500 000. We used the ΔK statistic (Evanno et al. 2005) to determine the most likely number of groups, and examined ancestry values for each individual and population graphically.

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