Distribution of sampling sites in the Iberian Peninsula
Black dots refer to populations of size > 19 individuals. Green background refers to elevation (from pale to dark green: low to high elevation).
Number of individuals in sampling areas across spatial scal
Mean: black; +/- SD: grey
Evolution of DeltaK among an increasing number of K (2 -20)
Sampling details of the 132 demes
(Long. and Lat. refer to geographic coordinates of sampling sites; N. is the number of individuals of demes; A. mean allelic richness; AR. corrected allelic richness, accounting to variation in deme size; Fis. Fis estimate of deme, computed without Mon01 and Mon 27).
Details of primer sequence and genotyping
Multiplexed PCR were performed in a 10 µL reaction volume using 25 ng of genomic DNA, 0.4 U of DreamTaq DNA Polymerase (Thermo Scientific®), 0.75 µL Dream Taq Green Buffer (including 20 mM MgCl2, Thermo Scientific®), 1 µM Betaine, 0.24 µL dNTP (10 µM) and deionized H2O. PCR amplifications were run on a Veriti® 96 well fast Thermal cycler (Applied Biosystems®) using the following settings: a first denaturation step at 95 °C during 10 min; 40 cycles of denaturation (30 s at 95 °C), hybridization (30 s at 55 °C) and elongation (1 min at 72 °C), and a final elongation step at 72 °C during 10 min. One µL of PCR products were denatured within a mix of 10 µL of formamide and 0.3 µL of 600 Liz marker before being run on an ABI PRISM 3500 sequencer (Life Technologies®). Genotypes were read using the software GENEMAPPER V 4.1 (Applied Biosystems®).