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Abbreviations indicate sampling locations see supplementary table S1Supplementary Material online for details.

Despite their recent divergence, D. In particular, laboratory crosses between D. During courtship, Drosophila males of many species produce species-specific courtship songs by vibrating their wings.

Differences in courtship song, especially in the interpulse interval IPI; the time from the end of a pulse to the start of the next have been shown to be important for premating isolation in several Drosophila species Saarikettu et al. Playback experiments of semispecies-specific songs increase the mating success of muted Old married women ready chat sex D.

Their geographic range and high degree of sexual isolation differentiate D. Thus, D.

However, despite the potential of D. Early studies in D.

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A restriction site survey of variation at a few nuclear loci found greater differentiation between the Wife want hot sex Pemberville semispecies at X-linked genes than at autosomal genes Ford and Aquadro However, beyond a handful of genes Yoon ; Ford et al. Here we utilize whole genome sequencing to study patterns of genomic differentiation in the Drosophila athabasca species complex.

We assemble a draft genome Women over 40 looking for sex bike Laramie conduct a whole-genome population analysis of D. In particular, we examine whether our genomic data support the behavioral and geographic stratification of individuals into three semispecies, and compare patterns of nucleotide diversity within and divergence between semispecies on the X chromosome versus autosomes.

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Historical demography leaves characteristic atures in the genome, and we use our population genomic data to conduct an analysis of current and ancestral population sizes, levels of gene flow, and the timing of population splits in the D. Finally, we discuss the potential of D. Methods Collection of Drosophila athabasca Flies Woman wants sex Christmas Michigan collected over banana bait during the summers of — To avoid creating artificial population structure as a result of sampling artifacts, we collected flies at 19 different locations widely spread across the D.

Over iso-female lines were established from these collection sites, and we used Sanger sequencing of a mitochondrial DNA fragment to confirm which lines belonged to the D. A total of D.

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Courtship Song Assays Courtship songs were recorded for 28 D. Both male and female virgins were collected shortly following eclosion and aged in individual vials for 7—10 days under the same temperature and lighting conditions as during rearing.

Recordings were captured by placing a single virgin male and virgin female in an Insectavox insect recording chamber Gorczyca and Hall Three separate mating pairs were recorded for each line, Adult want casual sex Dale City Virginia interpulse interval IPI from High-Repetition-Rate HRR song bursts was calculated directly from song waveforms as an average of the three pairs.

Genome Assembly and Annotation To create a reference genome assembly for D.

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We prepared a Bbw looking for more than a good time of four genomic libraries using standard Illumina protocols, two short insert paired-end libraries with mean insert sizes of 91 bp 24 sd and bp 63 sd from a genomic DNA extraction of 10 pooled females, and two additional mate-pair libraries with mean insert sizes of 2, bp sd and 4, bp sd from a genomic DNA extraction of 20 pooled females.

The two long-insert mate-pair libraries were cropped to 36 bp to reduce the chances of Private sex Athabasca over library construction breakpoints, as suggested by the manufacturer.

Re Private sex Athabasca screened and cropped for adapter and bacterial contamination, leaving a total of To aid in genome annotation, we made three mRNAseq libraries using the Granny fuck Winona md. Illumina mRNAseq libraries were prepared using standard protocols.

We sequenced each library from both ends for 76 bp on a lane of a GAII, resulting in 4.

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We then anchored the scaffolds onto chromosomes based on the D. We classified strains into semispecies groups based on Seeking Special Freind combination of geographic location and courtship song interpulse interval supplementary tables S1 and S2Supplementary Material online. Karyotype information was also collected following the method in Pimpinelli et al.

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Genomic DNA was extracted from a single female fly from each of the strains using the same method as. We sequenced 90 bp Avoca TX bi horny wives re, generating 2 Gb of sequence for each strain.

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Mean genomic coverage per strain was 9. Variants for each strain were called using the GATK pipeline [version 1.

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Local realignment was performed on the merged file around indel regions to prevent erroneous variant calls due to alignment error. Variants from all strains were called simultaneously. Due to the lack of validated SNPs in D.