WGS of Anemopaegma oligoneuron (Voucher Nogueira 171)
Total DNA was diluted to a final volume of 50 µl at 100 ng/ml and sonicated using Covaris S-series with the following parameters: 175W of incident power, 10% of duty factor, 200 cycles and 50 s. Fragmented sample was used to prepare pair-ended libraries using NEBNext DNA Library Prep Master Mix and NEBNext Multiplex oligos (New England Biolabs, Ipswich, USA), following the manufacturerÕs protocol with modifications proposed by M. Carlsen (pers. comm.). Library was quantified via qPCR using KAPA Library Quantification Kit for Illumina Sequencing Platforms (Kapa Biosystems, Wilmington, USA) on an ABI7300 RT-PCR. Sample was diluted in ddH2O to a final concentration of 2nM and 10 µl of diluted libraries were pooled together (21 samples total). The final pool was sequenced at the Laborat—rio Multiusu‡rios Centralizado (ESALq-USP, Piracicaba, Brazil). Fragment size: 250~300 bp. Read size: 101 bp.