title: Melittobia australica Ion Torrent PGM reads
design: Genomic DNA for the sequencing library was extracted from a pool of 20 females from a laboratory stock culture of M. australica (S line27) that was collected in Otsu, Shiga, Japan in 200015. DNA extraction was done using DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) according to the manufacturer instructions. DNA was eluted in 50 l of AE buffer and RNase I (Life Technologies, Carlsbad, CA, USA) was added at a concentration of 20 g/ml to digest the RNA. Genomic DNA library was constructed using Ion Xpress Plus Fragment Library Kit (Life Technologies, Carlsbad, CA, USA). After the fragmentation by the restriction enzyme and the adapter ligation, approximately 400 bp sized fragments (without the length of the adapters) were selected by gel excision (E-gel; Invitrogen, Carlsbad, CA, USA). To determine the appropriate dilution for the template preparation, the library was evaluated and quantified using a Bioanalyser 2100 (Agilent Technologies, Santa Clara, CA, USA). The diluted library was amplified by emulsion PCR using an Ion One Touch 2 (Life Technologies, Carlsbad, CA, USA) and enriched using an Ion One Touch ES (Life Technologies, Carlsbad, CA, USA) with an Ion PGM Hi-Q OT2 Kit (Life Technologies, Carlsbad, CA, USA). Sequencing was performed on an Ion Torrent PGM sequencer with an Ion PGM Hi-Q Sequencing Kit and Ion 318 Chip Kit v2 (Life Technologies, Carlsbad, CA, USA).