ddRADseq of Dailognatha hellenica: adult whole organism
Individual was killed and preserved in 100% ethanol and stored at -20C. Total genomic DNA was extracted from prothoracic or abdominal tissue using the DNeasy Blood and Tissue kit (Qiagen), following the manufacturers instructions. Double digest restriction-site associated DNA (ddRAD) sequencing library preparation followed the protocol of Peterson et al. (2012). In brief, lab work comprised of: (a) double-digestion of DNA with the restriction enzymes EcoRI and MseI, (b) ligation of unique barcodes (10 bp) and Illumina adapter sequences to the digested fragments, (c) pooling of the barcoded products into three libraries of 60-72 samples each, (d) size-selection of libraries for 350450 bp fragments using a Pippin Prep instrument (Sage Science Inc) and (e) PCR-amplification (10 cycles) of the size-selected fragments using high-fidelity DNA polymerase (iProof, Bio-Rad).
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