We selected a set of 21 microsatellite markers of which some were linked (located on the same chromosome) and some unlinked (situated on unique chromosomes) (Table S1) [38] , [42] (link), and genotyped a total of 206 blue tits. Summary statistics for all loci with their genomic location on the blue tit linkage map [30] (link) and the zebra finch genome assembly [35] (link) are given in Table S1. All birds were molecularly sexed by amplifying a Z- and W-linked locus, TGZ-002 (D. Dawson, University of Sheffield, unpublished), and this information was used for interpreting the genotypes of the Z-linked microsatellites.
All loci were PCR-amplified in three different multiplexes using QIAGEN Multiplex PCR kit (Qiagen, ltd.). Primer sequences and annealing temperatures for the microsatellites are given in Table S1 (see also Olano-Marin et al.[29] (link) and Hansson et al.[30] (link)). The PCR-products were separated and visualized using an ABI 3730 capillary sequencer (Applied Biosystems), and the genotypes were scored with Genemapper 4.0 (Applied Biosystems).
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