Data from: Double-digest RAD Sequencing using Ion Proton semiconductor platform (ddRADseq-ion) with non-model organisms
Data files
Mar 19, 2015 version files 22.09 MB
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Library1_SNPs.xlsx
3.81 MB
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Library10_SNPs.xlsx
1.16 MB
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Library11_SNPs.xlsx
1.66 MB
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Library12_SNPs.xlsx
757.85 KB
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Library13_SNPs.xlsx
409.63 KB
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Library2_SNPs.xlsx
984.01 KB
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Library3_SNPs.xlsx
1.76 MB
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Library4_SNPs.xlsx
2.38 MB
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Library5_SNPs.xlsx
1.64 MB
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Library6_SNPs.xlsx
1.11 MB
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Library7_SNPs.xlsx
1.45 MB
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Library8_SNPs.xlsx
2.25 MB
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Library9_SNPs.xlsx
1.31 MB
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RAxML_treefile.tre
284 B
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Replicate_1_genotypes_16x.raw
16.92 KB
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Replicate_1_genotypes_8x.raw
59.54 KB
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Replicate_2_genotypes_16x.raw
16.92 KB
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Replicate_2_genotypes_8x.raw
59.61 KB
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Table_1_Recknagel_et_al..xlsx
804.74 KB
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Table_2_Recknagel_et_al..xlsx
439.66 KB
Abstract
Research in evolutionary biology involving nonmodel organisms is rapidly shifting from using traditional molecular markers such as mtDNA and microsatellites to higher throughput SNP genotyping methodologies to address questions in population genetics, phylogenetics and genetic mapping. Restriction site associated DNA sequencing (RAD sequencing or RADseq) has become an established method for SNP genotyping on Illumina sequencing platforms. Here, we developed a protocol and adapters for double-digest RAD sequencing for Ion Torrent (Life Technologies; Ion Proton, Ion PGM) semiconductor sequencing. We sequenced thirteen genomic libraries of three different nonmodel vertebrate species on Ion Proton with PI chips: Arctic charr Salvelinus alpinus, European whitefish Coregonus lavaretus and common lizard Zootoca vivipara. This resulted in ~962 million single-end reads overall and a mean of ~74 million reads per library. We filtered the genomic data using Stacks, a bioinformatic tool to process RAD sequencing data. On average, we obtained ~11 000 polymorphic loci per library of 6–30 individuals. We validate our new method by technical and biological replication, by reconstructing phylogenetic relationships, and using a hybrid genetic cross to track genomic variants. Finally, we discuss the differences between using the different sequencing platforms in the context of RAD sequencing, assessing possible advantages and disadvantages. We show that our protocol can be used for Ion semiconductor sequencing platforms for the rapid and cost-effective generation of variable and reproducible genetic markers.