title: GBS of Caucasotachea vindobonensis and Dendrocopos syriacus/major
design: GBS libraries were prepared following previously described methodology (Elshire et al., 2011). The protocol included PstI restriction enzyme digestion and ligation of 48 indexed adapters. GBS libraries were prepared after few optimization steps, following previously described methodology (Elshire et al., 2011). The protocol included PstI restriction enzyme digestion and ligation of 48 indexed adapters designed with a GBS Barcode Generator software (http://www.deenabio.com/services/gbs-adapters). In brief, during library preparation process, commercially synthetized oligonucleotides (forward and reverse complement) containing unique barcodes (4-9 nt long) and common adapter were annealed to obtain ligation-ready double-stranded oligoes. Genomic DNA was digested overnight with tenfold PstI-HF enzyme excess. Digestion products were ligated with barcoded adapters using T4 DNA ligase. The obtained libraries were pooled and sequenced in single flowcell lane. The SRA data include .fastq file with multiplexed raw reads and ‘key’ file informing about samples and ligated barcodes. Within the reads the barcoded sequence is followed by PstI digestion site and random genomic sequences. Please use the ‘Woodpeckers_key.txt’ file to perform analysis in Dendrocopos sp. and ‘Snails_key.txt’ for analysis in Caucasotachea sp. when using GBS-dedicated software. Woodpeckers_key Flowcell Lane Barcode FullSampleName AC9H6DACXX 5 AACT DS1 AC9H6DACXX 5 CCAG DS2 AC9H6DACXX 5 TTGA DS3 AC9H6DACXX 5 GGTA DS4 AC9H6DACXX 5 ATTG DS5 AC9H6DACXX 5 CGGT DS6 AC9H6DACXX 5 TGCG DS8 AC9H6DACXX 5 GTAT DS9 AC9H6DACXX 5 AACCA DS11 AC9H6DACXX 5 CCACG DS12 AC9H6DACXX 5 TATAA DM1 AC9H6DACXX 5 GAGCG DM2 AC9H6DACXX 5 ACATA DM3 AC9H6DACXX 5 CTCAG DM4 AC9H6DACXX 5 TCTGG DM5 AC9H6DACXX 5 CGGCA DM6 AC9H6DACXX 5 GAATCA DM7 AC9H6DACXX 5 ACTGCA DM8 AC9H6DACXX 5 TTGCCA DM9 AC9H6DACXX 5 TGCACA DM10 AC9H6DACXX 5 GATCAA DM11 AC9H6DACXX 5 ACATGA DM12 Snails_key Flowcell Lane Barcode FullSampleName AC9H6DACXX 5 CTCGCA CV1 AC9H6DACXX 5 CTGAGA CV2 AC9H6DACXX 5 GGATACA CV3 AC9H6DACXX 5 AATGCCA CV4 AC9H6DACXX 5 TCGAACA CV5 AC9H6DACXX 5 TGCCGCA CV6 AC9H6DACXX 5 GACTCCA CV7 AC9H6DACXX 5 ACTAGCA CV8 AC9H6DACXX 5 CTAGACA CV9 AC9H6DACXX 5 AGGTCCA CV10 AC9H6DACXX 5 GAACAATA CV11 AC9H6DACXX 5 CCTACCGT CV12 AC9H6DACXX 5 TTCCTTAG CV13 AC9H6DACXX 5 AGGAGGCT CV14 AC9H6DACXX 5 GAAGACTG CV15 AC9H6DACXX 5 CCTCCTGA CV16 AC9H6DACXX 5 TTCTTACG CV17 AC9H6DACXX 5 CGGCGGAT CV18 AC9H6DACXX 5 GAAGAACCT CV19 AC9H6DACXX 5 ACCATCGCA CV20 AC9H6DACXX 5 TTGTCTAAT CV21 AC9H6DACXX 5 GGTTGGTCA CV22 AC9H6DACXX 5 AACGAACAT CV23 AC9H6DACXX 5 CCAACCTGA CV24 AC9H6DACXX 5 TTGCTTACA CV25 AC9H6DACXX 5 CGTGAGGTT CV26 